57 results on '"Tanya L. Daigle"'
Search Results
2. Local Connectivity and Synaptic Dynamics in Mouse and Human Neocortex
- Author
-
Luke Campagnola, Stephanie C. Seeman, Thomas Chartrand, Lisa Kim, Alex Hoggarth, Clare Gamlin, Shinya Ito, Jessica Trinh, Pasha Davoudian, Cristina Radaelli, Mean-Hwan Kim, Travis Hage, Thomas Braun, Lauren Alfiler, Julia Andrade, Phillip Bohn, Rachel Dalley, Alex Henry, Sara Kebede, Mukora Alice, David Sandman, Grace Williams, Rachael Larsen, Corinne Teeter, Tanya L. Daigle, Kyla Berry, Nadia Dotson, Rachel Enstrom, Melissa Gorham, Madie Hupp, Samuel Dingman Lee, Kiet Ngo, Philip R. Nicovich, Lydia Potekhina, Shea Ransford, Amanda Gary, Jeff Goldy, Delissa McMillen, Trangthanh Pham, Michael Tieu, La’Akea Siverts, Miranda Walker, Colin Farrell, Martin Schroedter, Cliff Slaughterbeck, Charles Cobb, Richard Ellenbogen, Ryder P. Gwinn, C. Dirk Keene, Andrew L. Ko, Jeffrey G. Ojemann, Daniel L. Silbergeld, Daniel Carey, Tamara Casper, Kirsten Crichton, Michael Clark, Nick Dee, Lauren Ellingwood, Jessica Gloe, Matthew Kroll, Josef Sulc, Herman Tung, Katherine Wadhwani, Krissy Brouner, Tom Egdorf, Michelle Maxwell, Medea McGraw, Christina Alice Pom, Augustin Ruiz, Jasmine Bomben, David Feng, Nika Hejazinia, Shu Shi, Aaron Szafer, Wayne Wakeman, John Phillips, Amy Bernard, Luke Esposito, Florence D. D’Orazi, Susan Sunkin, Kimberly Smith, Bosiljka Tasic, Anton Arkhipov, Staci Sorensen, Ed Lein, Christof Koch, Gabe Murphy, Hongkui Zeng, and Tim Jarsky
- Subjects
Adult ,Male ,Neurons ,Multidisciplinary ,Models, Neurological ,Datasets as Topic ,Excitatory Postsynaptic Potentials ,Mice, Transgenic ,Neocortex ,Synaptic Transmission ,Article ,Temporal Lobe ,Mice ,Inhibitory Postsynaptic Potentials ,Neural Pathways ,Synapses ,Animals ,Humans ,Female ,Visual Cortex - Abstract
We present a unique, extensive, and open synaptic physiology analysis platform and dataset. Through its application, we reveal principles that relate cell type to synaptic properties and intralaminar circuit organization in the mouse and human cortex. The dynamics of excitatory synapses align with the postsynaptic cell subclass, whereas inhibitory synapse dynamics partly align with presynaptic cell subclass but with considerable overlap. Synaptic properties are heterogeneous in most subclass-to-subclass connections. The two main axes of heterogeneity are strength and variability. Cell subclasses divide along the variability axis, whereas the strength axis accounts for substantial heterogeneity within the subclass. In the human cortex, excitatory-to-excitatory synaptic dynamics are distinct from those in the mouse cortex and vary with depth across layers 2 and 3.
- Published
- 2022
3. RecV recombinase system for in vivo targeted optogenomic modifications of single cells or cell populations
- Author
-
Mark J. Schnitzer, Anat Kahan, Andrew Curtright, Qingming Luo, Yun Wang, Bosiljka Tasic, Ajay Dhaka, Ali Cetin, Hui Gong, Shenqin Yao, Marty Mortrud, Xiuli Kuang, Tanya L. Daigle, Shaoqun Zeng, Soumya Chatterjee, Radosław Chrapkiewicz, Peng Yuan, Viviana Gradinaru, Pooja Balaram, Thomas Zhou, Hongkui Zeng, and Ben Ouellette
- Subjects
Flp ,Computational biology ,Biology ,Optogenetics ,Biochemistry ,Genome ,Article ,recombinase ,Recombinases ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,RecV ,Recombinase ,Biological neural network ,Animals ,Molecular Biology ,Zebrafish ,030304 developmental biology ,optogenomic ,Neurons ,Regulation of gene expression ,0303 health sciences ,Brain ,Cre ,Dre ,Genomics ,Cell Biology ,biology.organism_classification ,recombination ,Vivid ,Gene Expression Regulation ,chemistry ,Light-inducible ,Genetic Engineering ,DNA ,Function (biology) ,Biotechnology - Abstract
Brain circuits comprise vast numbers of intricately interconnected neurons with diverse molecular, anatomical and physiological properties. To allow “user-defined” targeting of individual neurons for structural and functional studies, we created light-inducible site-specific DNA recombinases (SSRs) based on Cre, Dre and Flp (RecVs). RecVs can induce genomic modifications by one-photon or two-photon light induction in vivo. They can produce targeted, sparse and strong labeling of individual neurons by modifying multiple loci within mouse and zebrafish genomes. In combination with other genetic strategies, they allow intersectional targeting of different neuronal classes. In the mouse cortex they enable sparse labeling and whole-brain morphological reconstructions of individual neurons. Furthermore, these enzymes allow single-cell two-photon targeted genetic modifications and can be used in combination with functional optical indicators with minimal interference. In summary, RecVs enable spatiotemporally-precise optogenomic modifications that can facilitate detailed single-cell analysis of neural circuits by linking genetic identity, morphology, connectivity and function.
- Published
- 2020
4. Long-term labeling and imaging of synaptically-connected neuronal networksin vivousing double-deletion-mutant rabies viruses
- Author
-
Lei Jin, Heather A. Sullivan, Mulangma Zhu, Thomas K. Lavin, Makoto Matsuyama, Xin Fu, Nicholas E. Lea, Ran Xu, YuanYuan Hou, Luca Rutigliani, Maxwell Pruner, Kelsey R. Babcock, Jacque Pak Kan Ip, Ming Hu, Tanya L. Daigle, Hongkui Zeng, Mriganka Sur, Guoping Feng, and Ian R. Wickersham
- Abstract
Monosynaptic tracing is a widely-used technique for mapping neural circuitry, but its cytotoxicity has confined it primarily to anatomical applications. Here we present a second-generation system for labeling direct inputs to targeted neuronal populations with minimal toxicity, using double-deletion-mutant rabies viruses. Spread of the viruses requires expression of both deleted viral genesin transin postsynaptic source cells; suppressing this expression with doxycycline following an initial period of viral replication reduces toxicity to postsynaptic cells. Longitudinal two-photon imagingin vivoindicated that over 90% of both presynaptic and source cells survived for the full twelve-week course of imaging.Ex vivowhole-cell recordings at 5 weeks postinfection showed that the second-generation system perturbs input and source cells much less than does the first-generation system. Finally, two-photon calcium imaging of labeled networks of visual cortex neurons showed that their visual response properties appeared normal for 10 weeks, the longest we followed them.
- Published
- 2021
5. Comparative cellular analysis of motor cortex in human, marmoset and mouse
- Author
-
Owen White, Kimberly A. Smith, Brian D. Aevermann, William J. Romanow, Joseph R. Ecker, Michael Tieu, Michael Hawrylycz, Sheng-Yong Niu, Brian R. Herb, Jacinta Lucero, Sten Linnarsson, Tanya L. Daigle, Christine S. Liu, Ed S. Lein, Boudewijn P. F. Lelieveldt, Zizhen Yao, Yang Eric Li, Stephan Fischer, Trygve E. Bakken, Jeremy A. Miller, C. Dirk Keene, Scott F. Owen, Wei Tian, Joshua Orvis, Nongluk Plongthongkum, Rosa Castanon, Megan Crow, Thomas Höllt, Bing Ren, Darren Bertagnolli, Weixiu Dong, Herman Tung, Baldur van Lew, Delissa McMillen, Bosiljka Tasic, Angeline Rivkin, Eran A. Mukamel, Nora Reed, Alexander Dobin, Chongyuan Luo, Patrick R. Hof, Nick Dee, Rongxin Fang, Kirsten Crichton, M. Margarita Behrens, Anna Bartlett, Renee Zhang, Olivier Poirion, Josef Sulc, Philip R. Nicovich, Rebecca D. Hodge, Evan Z. Macosko, Staci A. Sorensen, Dinh Diep, Thanh Pham, Songlin Ding, Richard H. Scheuermann, Jayaram Kancherla, Jeroen Eggermont, Seth A. Ament, Ronna Hertzano, Jeff Goldy, Christine Rimorin, Julia K. Osteen, Kimberly Siletti, Steven A. McCarroll, Hanqing Liu, C. Palmer, Saroja Somasundaram, Jonathan T. Ting, Jerold Chun, Xiaomeng Hou, Guoping Feng, Kun Zhang, Fenna M. Krienen, Blue B. Lake, Amy Torkelson, Hongkui Zeng, Sebastian Preissl, Christof Koch, Nikolas L. Jorstad, Andrew L. Ko, Héctor Corrada Bravo, Aviv Regev, Nikolai C. Dembrow, Kanan Lathia, Antonio Pinto-Duarte, Xinxin Wang, Lucas T. Graybuck, Melissa Goldman, Marmar Moussa, William J. Spain, Peter V. Kharchenko, Qiwen Hu, Adriana E. Sedeno-Cortes, Gregory D. Horwitz, Rachel A. Dalley, Anup Mahurkar, Brian E. Kalmbach, Andrew Aldridge, Jesse Gillis, Anna Marie Yanny, Joseph R. Nery, Tamara Casper, Fangming Xie, and Matthew Kroll
- Subjects
Epigenomics ,Male ,Cell type ,Genetics of the nervous system ,Computational biology ,Biology ,Molecular neuroscience ,Article ,Epigenesis, Genetic ,Transcriptome ,Mice ,Atlases as Topic ,Glutamates ,Species Specificity ,Molecular evolution ,Animals ,Humans ,GABAergic Neurons ,Gene ,In Situ Hybridization, Fluorescence ,Phylogeny ,Neurons ,Multidisciplinary ,Gene Expression Profiling ,Motor Cortex ,Callithrix ,Epigenome ,Middle Aged ,Cellular neuroscience ,Chromatin ,Organ Specificity ,DNA methylation ,Female ,Single-Cell Analysis - Abstract
The primary motor cortex (M1) is essential for voluntary fine-motor control and is functionally conserved across mammals1. Here, using high-throughput transcriptomic and epigenomic profiling of more than 450,000 single nuclei in humans, marmoset monkeys and mice, we demonstrate a broadly conserved cellular makeup of this region, with similarities that mirror evolutionary distance and are consistent between the transcriptome and epigenome. The core conserved molecular identities of neuronal and non-neuronal cell types allow us to generate a cross-species consensus classification of cell types, and to infer conserved properties of cell types across species. Despite the overall conservation, however, many species-dependent specializations are apparent, including differences in cell-type proportions, gene expression, DNA methylation and chromatin state. Few cell-type marker genes are conserved across species, revealing a short list of candidate genes and regulatory mechanisms that are responsible for conserved features of homologous cell types, such as the GABAergic chandelier cells. This consensus transcriptomic classification allows us to use patch–seq (a combination of whole-cell patch-clamp recordings, RNA sequencing and morphological characterization) to identify corticospinal Betz cells from layer 5 in non-human primates and humans, and to characterize their highly specialized physiology and anatomy. These findings highlight the robust molecular underpinnings of cell-type diversity in M1 across mammals, and point to the genes and regulatory pathways responsible for the functional identity of cell types and their species-specific adaptations., An examination of motor cortex in humans, marmosets and mice reveals a generally conserved cellular makeup that is likely to extend to many mammalian species, but also differences in gene expression, DNA methylation and chromatin state that lead to species-dependent specializations.
- Published
- 2021
6. Ultra-precise all-optical manipulation of neural circuits with multifunctional Cre-dependent transgenic mice
- Author
-
Masato Sadahiro, Hayley A. Bounds, Ian A. Oldenburg, Karthika Gopakumar, Tanya L. Daigle, Marta Gajowa, William D. Hendricks, Daniel Quintana, Hongkui Zeng, and Hillel Adesnik
- Subjects
Nervous system ,Opsin ,Calcium imaging ,medicine.anatomical_structure ,Neural ensemble ,Computer science ,GCaMP ,medicine ,Biological neural network ,Optogenetics ,Neuroscience ,Photostimulation - Abstract
SummaryDetermining which features of the neural code drive perception and behavior requires the ability to simultaneous read out and write in neural activity patterns with high precision across many neurons. All-optical systems that combine two photon (2p) calcium imaging and targeted 2p photostimulation enable the activation of specific, functionally defined groups of neurons in behaving animals. However, these techniques do not yet have the ability to reveal how the specific distribution of firing rates across a relevant neural population mediates neural computation and behavior. The key technical obstacle is the inability to transform single-cell calcium signals into accurate estimates of firing rate changes and then write in these cell-specific firing rate changes to each individual neuron in a targeted population. To overcome this challenge, we made two advances: first we introduce a new genetic line of mice for robust Cre-dependent co-expression of a high-performance calcium indicator and a potent soma-targeted microbial opsin. Second, using this line, we developed a pipeline that enables the read-out and write-in of precise population vectors of neural activity across a targeted group of neurons. The combination of the new multifunctional transgenic line and the photostimulation paradigm offer a powerful and convenient platform for investigating the neural codes of computation and behavior. It may prove particularly useful for probing causal features of the geometry of neural representations where the ability to directly control the topology of population activity is essential.
- Published
- 2021
7. A whole-brain monosynaptic input connectome to neuron classes in mouse visual cortex
- Author
-
Kat North, Jasmin Bomben, Shenqin Yao, Bosiljka Tasic, Nicole Hancock, Shea Ransford, Thuyanh V. Nguyen, Ali Williford, Karla E. Hirokawa, Wayne Wakeman, Stefan Mihalas, Lydia Ng, Fiona Griffin, Robert Howard, Julie A. Harris, Rachel Enstrom, Tom Egdorf, Nadezhda Dotson, Eric Lee, Kyla Mace, Amanda Gary, Peter A. Groblewski, Michelle Maxwell, Lydia Potekhina, Tanya L. Daigle, Nhan-Kiet Ngo, Medea McGraw, Krissy Brouner, Philip R. Nicovich, Benjamin Ouellette, Jennifer Luviano, Emily Gelfand, Sam Seid, Chelsea Nayan, Maitham Naeemi, Marty Mortrud, Jackie Swapp, Cho A, Ali Cetin, Hong Gu, Hongkui Zeng, M. J. Taormina, Sophie Lambert, Leonard Kuan, Ruweida Ahmed, Thomas Zhou, Melissa Gorham, Augustin Ruiz, Linzy Casal, Shiella Caldejon, and Quanxin Wang
- Subjects
General Neuroscience ,Rabies virus ,Tracing ,Biology ,medicine.disease_cause ,Entire visual cortex ,Visual cortex ,medicine.anatomical_structure ,medicine ,Connectome ,Excitatory postsynaptic potential ,Neuron ,Neuroscience ,Brain function - Abstract
Identification of the structural connections between neurons is a prerequisite to understanding brain function. We developed a pipeline to systematically map brain-wide monosynaptic inputs to specific neuronal populations using Cre-driver mouse lines and the recombinant rabies tracing system. We first improved the rabies virus tracing strategy to accurately identify starter cells and to efficiently quantify presynaptic inputs. We then mapped brain-wide presynaptic inputs to different excitatory and inhibitory neuron subclasses in the primary visual cortex and seven higher visual areas. Our results reveal quantitative target-, layer- and cell-class-specific differences in the retrograde connectomes, despite similar global input patterns to different neuronal populations in the same anatomical area. The retrograde connectivity we define is consistent with the presence of the ventral and dorsal visual information processing streams and reveals further subnetworks within the dorsal stream. The hierarchical organization of the entire visual cortex can be derived from intracortical feedforward and feedback pathways mediated by upper- and lower-layer input neurons, respectively. This study expands our knowledge of the brain-wide inputs regulating visual areas and demonstrates that our improved rabies virus tracing strategy can be used to scale up the effort in dissecting connectivity of genetically defined cell populations in the whole mouse brain.
- Published
- 2021
8. Intersectional mapping of multi-transmitter neurons and other cell types in the brain
- Author
-
Jian Xu, Andrew Jo, Raina P. DeVries, Sercan Deniz, Suraj Cherian, Idris Sunmola, Xingqi Song, John J. Marshall, Katherine A. Gruner, Tanya L. Daigle, Anis Contractor, Talia N. Lerner, Hongkui Zeng, and Yongling Zhu
- Subjects
Neurons ,Habenula ,Mice ,Vesicular Glutamate Transport Proteins ,Animals ,Mice, Transgenic ,General Biochemistry, Genetics and Molecular Biology - Abstract
Recent developments in intersectional strategies have greatly advanced our ability to precisely target brain cell types based on unique co-expression patterns. To accelerate the application of intersectional genetics, we perform a brain-wide characterization of 13 Flp and tTA mouse driver lines and selected seven for further analysis based on expression of vesicular neurotransmitter transporters. Using selective Cre driver lines, we created more than 10 Cre/tTA combinational lines for cell type targeting and circuit analysis. We then used VGLUT-Cre/VGAT-Flp combinational lines to identify and map 30 brain regions containing neurons that co-express vesicular glutamate and gamma-aminobutyric acid (GABA) transporters, followed by tracing their projections with intersectional viral vectors. Focusing on the lateral habenula (LHb) as a target, we identified glutamatergic, GABAergic, or co-glutamatergic/GABAergic innervations from ∼40 brain regions. These data provide an important resource for the future application of intersectional strategies and expand our understanding of the neuronal subtypes in the brain.
- Published
- 2021
9. Motion/direction-sensitive thalamic neurons project extensively to the middle layers of primary visual cortex
- Author
-
Tanya L. Daigle, Yun Wang, Bosiljka Tasic, Jun Zhuang, R. Clay Reid, Hongkui Zeng, Emily Turschak, Kevin T. Takasaki, Jack Waters, Naveen D. Ouellette, and Rylan S. Larsen
- Subjects
Visual cortex ,medicine.anatomical_structure ,nervous system ,Population level ,Middle layer ,media_common.quotation_subject ,Thalamus ,medicine ,Motion direction ,Contrast (vision) ,Biology ,Neuroscience ,media_common - Abstract
The motion/direction-sensitive and location-sensitive neurons are two major functional types in mouse visual thalamus that project to the primary visual cortex (V1). It has been proposed that the motion/direction-sensitive neurons mainly target the superficial layers in V1, in contrast to the location-sensitive neurons which mainly target the middle layers. Here, by imaging calcium activities of motion/direction-sensitive and location-sensitive axons in V1, we find no evidence for these cell-type specific laminar biases at population level. Furthermore, using a novel approach to reconstruct single-axon structures with identified in vivo response types, we show that, at single-axon level, the motion/direction-sensitive axons have middle layer preferences and project more densely to the middle layers than the location-sensitive axons. Overall, our results demonstrate that Motion/direction-sensitive thalamic neurons project extensively to the middle layers of V1, challenging the current view of the thalamocortical organizations in the mouse visual system.
- Published
- 2021
10. Laminar distribution and arbor density of two functional classes of thalamic inputs to primary visual cortex
- Author
-
Emily Turschak, Yun Wang, Bosiljka Tasic, Kevin T. Takasaki, R. Clay Reid, Jun Zhuang, Hongkui Zeng, Rylan S. Larsen, Jack Waters, Tanya L. Daigle, and Naveen D. Ouellette
- Subjects
Male ,Cell type ,Population ,Thalamus ,Motion Perception ,Mice, Transgenic ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Article ,Calcium imaging ,Orientation ,Primary Visual Cortex ,medicine ,Animals ,Visual Pathways ,Calcium Signaling ,Axon ,Projection (set theory) ,education ,education.field_of_study ,Microscopy, Confocal ,Laminar flow ,Axons ,medicine.anatomical_structure ,Visual cortex ,Microscopy, Fluorescence, Multiphoton ,nervous system ,Female ,Neuroscience ,Photic Stimulation - Abstract
SUMMARY Motion/direction-sensitive and location-sensitive neurons are the two major functional types in mouse visual thalamus that project to the primary visual cortex (V1). It is under debate whether motion/direction-sensitive inputs preferentially target the superficial layers in V1, as opposed to the location-sensitive inputs, which preferentially target the middle layers. Here, by using calcium imaging to measure the activity of motion/direction-sensitive and location-sensitive axons in V1, we find evidence against these cell-type-specific laminar biases at the population level. Furthermore, using an approach to reconstruct axon arbors with identified in vivo response types, we show that, at the single-axon level, the motion/direction-sensitive axons project more densely to the middle layers than the location-sensitive axons. Overall, our results demonstrate that motion/direction-sensitive thalamic neurons project extensively to the middle layers of V1 at both the population and single-cell levels, providing further insight into the organization of thalamocortical projection in the mouse visual system., In brief Zhuang et al., investigate the functionally specific thalamocortical projection patterns in mouse primary visual cortex at both the population and single-axon levels. They find that the motion/direction-sensitive axons project extensively to the middle layers of primary visual cortex, challenging an existing hypothesis proposing a superficial targeting bias of these axons., Graphical Abstract
- Published
- 2021
11. Local Connectivity and Synaptic Dynamics in Mouse and Human Neocortex
- Author
-
Michael Tieu, Amy Bernard, Lisa Kim, Samuel Dingman Lee, Tim Jarsky, Corinne Teeter, Martin Schroedter, Alex Hoggarth, Kimberly A. Smith, Amanda Gary, Charles Cobb, John W. Phillips, Christina A. Pom, Herman Tung, Hongkui Zeng, Daniel Carey, Phillip Bohn, Colin Farrell, Bosiljka Tasic, Rusty Nicovich, Medea McGraw, Krissy Brouner, Andrew L. Ko, Katherine Wadhwani, Lauren Ellingwood, Tom Egdorf, Anton Arkhipov, Aaron Szafer, Michael Clark, Kirsten Crichton, Kyla Berry, Josef Sulc, Nick Dee, Gabe J. Murphy, Luke Esposito, Trangthanh Pham, Thomas Chartrand, Alex M. Henry, Rachel A. Dalley, Rachel Enstrom, Thomas Braun, Luke Campagnola, Cristina Radaelli, C. Dirk Keene, Tanya L. Daigle, Cliff Slaughterbeck, Sara Kebede, Rachael Larsen, Jeffrey G. Ojemann, Juia Andrade, Michelle Maxwell, Staci A. Sorensen, Jeff Goldy, Jessica Gloe, David Sandman, Shinya Ito, Susan M. Sunkin, Wayne Wakeman, Travis A. Hage, Melissa Gorham, Ryder P. Gwinn, Pasha A. Davoudian, Augustin Ruiz, Grace Williams, Clare Gamlin, Christof Koch, La'Akea Siverts, Stephanie C. Seeman, Jasmine Bomben, Florence D’Orazi, Madie Hupp, Ed S. Lein, Nadia Dotson, Shea Ransford, Nika Hejazinia, Mean Hwan Kim, Delissa McMillen, David Feng, Jessica Trinh, Lydia Potekhina, Alice Mukora, Lauren Alfiler, Tamara Casper, Shu Shi, Matthew Kroll, Kiet Ngo, Richard G. Ellenbogen, Daniel L. Silbergeld, and Miranda Walker
- Subjects
Synapse ,Cell type ,Neocortex ,medicine.anatomical_structure ,Excitatory synapse ,Cortex (anatomy) ,medicine ,Excitatory postsynaptic potential ,Biology ,Inhibitory postsynaptic potential ,Neuroscience ,Subclass - Abstract
To elucidate cortical microcircuit structure and synaptic properties we present a unique, extensive, and public synaptic physiology dataset and analysis platform. Through its application, we reveal principles that relate cell type to synapse properties and intralaminar circuit organization in the mouse and human cortex. The dynamics of excitatory synapses align with the postsynaptic cell subclass, whereas inhibitory synapse dynamics partly align with presynaptic cell subclass but with considerable overlap. Despite these associations, synaptic properties are heterogeneous in most subclass to subclass connections. The two main axes of heterogeneity are strength and variability. Cell subclasses divide along the variability axis, while the strength axis accounts for significant heterogeneity within the subclass. In human cortex, excitatory to excitatory synapse dynamics are distinct from those in mouse and short-term plasticity varies with depth across layers 2 and 3. With a novel connectivity analysis that enables fair comparisons between circuit elements, we find that intralaminar connection probability among cell subclasses exhibits a strong layer dependence.These and other findings combined with the analysis platform create new opportunities for the neuroscience community to advance our understanding of cortical microcircuits.
- Published
- 2021
12. Classification of electrophysiological and morphological neuron types in the mouse visual cortex
- Author
-
David Sandman, Brian Lee, Michael Hawrylycz, Sara Kebede, Tom Egdorf, David Reid, Rob Young, Nivretta Thatra, Stefan Mihalas, David Feng, John W. Phillips, Rebecca de Frates, DiJon Hill, Cliff Slaughterbeck, Samuel R Josephsen, Tamara Casper, Xiaoxiao Liu, Hanchuan Peng, Peter Chong, Colin Farrell, Zhi Zhou, Sheana Parry, Jed Perkins, Brian Long, Susan M. Sunkin, Matthew Kroll, Krissy Brouner, Melissa Gorham, Aaron Szafer, Wayne Wakeman, Hong Gu, Marissa Garwood, Daniel Park, Kristen Hadley, Michael S. Fisher, Lydia Potekhina, Ed Lein, Alice Mukora, Hongkui Zeng, Nick Dee, Aaron Oldre, Lindsay Ng, Thomas Braun, Grace Williams, Tracy Lemon, Julie A. Harris, Medea McGraw, Nadezhda Dotson, Philip R. Nicovich, Amanda Gary, Rusty Mann, Alex M. Henry, Caroline Habel, Samuel Dingman, Katherine E. Link, Nathalie Gaudreault, Gilberto J. Soler-Llavina, Thuc Nghi Nguyen, Nicole Blesie, Bosiljka Tasic, Lydia Ng, Christine Cuhaciyan, Tim Jarsky, Keith B. Godfrey, Costas A. Anastassiou, Kirsten Crichton, Josef Sulc, Martin Schroedter, Dan Castelli, Miranda Robertson, Amy Bernard, Lisa Kim, Songlin Ding, Alyse Doperalski, Nathan W. Gouwens, Herman Tung, Tsega Desta, Corinne Teeter, James Harrington, Jonathan T. Ting, Kris Bickley, Anton Arkhipov, Kiet Ngo, Changkyu Lee, Jim Berg, Agata Budzillo, Emma Garren, Tanya L. Daigle, Christof Koch, Rachel A. Dalley, Eliza Barkan, Staci A. Sorensen, Gabe J. Murphy, Shiella Caldejon, and Naz Taskin
- Subjects
0301 basic medicine ,Genetically modified mouse ,Cell type ,Patch-Clamp Techniques ,Databases, Factual ,Action Potentials ,Datasets as Topic ,Mice, Transgenic ,Biology ,Article ,Neuron types ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Genes, Reporter ,Biocytin ,medicine ,Animals ,Cell shape ,Cell Shape ,Visual Cortex ,Neurons ,General Neuroscience ,Laboratory mouse ,Electrophysiology ,030104 developmental biology ,Visual cortex ,medicine.anatomical_structure ,chemistry ,Transcriptome ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Understanding the diversity of cell types in the brain has been an enduring challenge and requires detailed characterization of individual neurons in multiple dimensions. To systematically profile morpho-electric properties of mammalian neurons, we established a single-cell characterization pipeline using standardized patch-clamp recordings in brain slices and biocytin-based neuronal reconstructions. We built a publicly accessible online database, the Allen Cell Types Database, to display these datasets. Intrinsic physiological properties were measured from 1,938 neurons from the adult laboratory mouse visual cortex, morphological properties were measured from 461 reconstructed neurons, and 452 neurons had both measurements available. Quantitative features were used to classify neurons into distinct types using unsupervised methods. We established a taxonomy of morphologically and electrophysiologically defined cell types for this region of the cortex, with 17 electrophysiological types, 38 morphological types and 46 morpho-electric types. There was good correspondence with previously defined transcriptomic cell types and subclasses using the same transgenic mouse lines.
- Published
- 2019
13. Signature morpho-electric, transcriptomic, and dendritic properties of extratelencephalic-projecting human layer 5 neocortical pyramidal neurons
- Author
-
Tanya L. Daigle, Cristina Radaelli, Scott F. Owen, Andrew L. Ko, Ryder P. Gwinn, Anna Marie Yanny, Kimberly A. Smith, Christopher Dirk Keene, Rachel A. Dalley, Medea McGraw, Ed S. Lein, Jonathan T. Ting, Brian E. Kalmbach, Hongkui Zeng, Anoop P. Patel, de Frates R, Jeffrey G. Ojemann, Matthew Mallory, Philip R. Nicovich, Richard G. Ellenbogen, Staci A. Sorensen, Daniel L. Silbergeld, Rebecca D. Hodge, Lucas T. Graybuck, Nick Dee, Nikolas L. Jorstad, Christof Koch, Trygve E. Bakken, Charles Cobbs, and Bosiljka Tasic
- Subjects
Cell type ,Neocortex ,Rodent ,biology ,Functional specialization ,Cell ,Phenotype ,Transcriptome ,medicine.anatomical_structure ,nervous system ,biology.animal ,medicine ,Neuroscience ,Function (biology) - Abstract
In the neocortex, subcerebral axonal projections originate largely from layer 5 (L5) extratelencephalic-projecting (ET) neurons. The highly distinctive morpho-electric properties of these neurons have mainly been described in rodents, where ET neurons can be labeled by retrograde tracers or transgenic lines. Similar labeling strategies are not possible in the human neocortex, rendering the translational relevance of findings in rodents unclear. We leveraged the recent discovery of a transcriptomically-defined L5 ET neuron type to study the properties of human L5 ET neurons in neocortical brain slices derived from neurosurgeries. Patch-seq recordings, where transcriptome, physiology and morphology are assayed from the same cell, revealed many conserved morpho-electric properties of human and rodent L5 ET neurons. Divergent properties were also apparent but were often smaller than differences between cell types within these two species. These data suggest a conserved function of L5 ET neurons in the neocortical hierarchy, but also highlight marked phenotypic divergence possibly related to functional specialization of human neocortex.
- Published
- 2020
14. Brain-wide single neuron reconstruction reveals morphological diversity in molecularly defined striatal, thalamic, cortical and claustral neuron types
- Author
-
Susan M. Sunkin, Zizhen Yao, Qi Li, Tanya L. Daigle, Yun Wang, Michael Hawrylycz, Jia Yuan, Donghuan Lu, Bosiljka Tasic, Lulu Yin, Yuanyuan Song, Z. Josh Huang, Karla E. Hirokawa, Zheng Yefeng, Matthew B. Veldman, Lei Huang, Luke Esposito, Feng Xiong, Shaoqun Zeng, An Liu, Liya Ding, Guodong Hong, Jintao Pan, Yaoyao Li, Wei Xiong, Qiang Ouyang, Yang Yu, Thuc Nghi Nguyen, Qingming Luo, Yimin Wang, Xiangning Li, Mengya Chen, Tao Wang, Zhangcan Ding, Lei Qu, Lydia Ng, Min Ye, Hsien-Chi Kuo, Peng Xie, Yuanyuan Li, Rachael Larsen, Zhixi Yun, Chris Hill, Julie A. Harris, Peng Wang, Longfei Li, Elise Shen, Lijuan Liu, Wan Wan, Sujun Zhao, Hui Gong, Zhongze Gu, Zongcai Ruan, Jing Yuan, Christof Koch, Xiangdong Yang, Wenjie Xu, Hongkui Zeng, Aaron Feiner, Stephanie Mok, Yanjun Duan, Shichen Zhang, Chao Chen, Yaping Wang, Wayne Wakeman, Phil Lesnar, Sara Kebede, Ping He, Staci A. Sorensen, Zijun Zhao, Anan Li, Hanchuan Peng, Xiuli Kuang, Shengdian Jiang, Zhi Zhou, Quanxin Wang, and Wei Xie
- Subjects
Cell type ,medicine.anatomical_structure ,nervous system ,Cortex (anatomy) ,Thalamus ,medicine ,Striatum ,Neuron ,Biology ,Axon ,Projection (set theory) ,Claustrum ,Neuroscience - Abstract
Ever since the seminal findings of Ramon y Cajal, dendritic and axonal morphology has been recognized as a defining feature of neuronal types. Yet our knowledge concerning the diversity of neuronal morphologies, in particular distal axonal projection patterns, is extremely limited. To systematically obtain single neuron full morphology on a brain-wide scale, we established a platform with five major components: sparse labeling, whole-brain imaging, reconstruction, registration, and classification. We achieved sparse, robust and consistent fluorescent labeling of a wide range of neuronal types by combining transgenic or viral Cre delivery with novel transgenic reporter lines. We acquired high-resolution whole-brain fluorescent images from a large set of sparsely labeled brains using fluorescence micro-optical sectioning tomography (fMOST). We developed a set of software tools for efficient large-volume image data processing, registration to the Allen Mouse Brain Common Coordinate Framework (CCF), and computer-assisted morphological reconstruction. We reconstructed and analyzed the complete morphologies of 1,708 neurons from the striatum, thalamus, cortex and claustrum. Finally, we classified these cells into multiple morphological and projection types and identified a set of region-specific organizational rules of long-range axonal projections at the single cell level. Specifically, different neuron types from different regions follow highly distinct rules in convergent or divergent projection, feedforward or feedback axon termination patterns, and between-cell homogeneity or heterogeneity. Major molecularly defined classes or types of neurons have correspondingly distinct morphological and projection patterns, however, we also identify further remarkably extensive morphological and projection diversity at more fine-grained levels within the major types that cannot presently be accounted for by preexisting transcriptomic subtypes. These insights reinforce the importance of full morphological characterization of brain cell types and suggest a plethora of ways different cell types and individual neurons may contribute to the function of their respective circuits.
- Published
- 2020
15. Author Correction: Comparative cellular analysis of motor cortex in human, marmoset and mouse
- Author
-
Trygve E. Bakken, Nikolas L. Jorstad, Qiwen Hu, Blue B. Lake, Wei Tian, Brian E. Kalmbach, Megan Crow, Rebecca D. Hodge, Fenna M. Krienen, Staci A. Sorensen, Jeroen Eggermont, Zizhen Yao, Brian D. Aevermann, Andrew I. Aldridge, Anna Bartlett, Darren Bertagnolli, Tamara Casper, Rosa G. Castanon, Kirsten Crichton, Tanya L. Daigle, Rachel Dalley, Nick Dee, Nikolai Dembrow, Dinh Diep, Song-Lin Ding, Weixiu Dong, Rongxin Fang, Stephan Fischer, Melissa Goldman, Jeff Goldy, Lucas T. Graybuck, Brian R. Herb, Xiaomeng Hou, Jayaram Kancherla, Matthew Kroll, Kanan Lathia, Baldur van Lew, Yang Eric Li, Christine S. Liu, Hanqing Liu, Jacinta D. Lucero, Anup Mahurkar, Delissa McMillen, Jeremy A. Miller, Marmar Moussa, Joseph R. Nery, Philip R. Nicovich, Sheng-Yong Niu, Joshua Orvis, Julia K. Osteen, Scott Owen, Carter R. Palmer, Thanh Pham, Nongluk Plongthongkum, Olivier Poirion, Nora M. Reed, Christine Rimorin, Angeline Rivkin, William J. Romanow, Adriana E. Sedeño-Cortés, Kimberly Siletti, Saroja Somasundaram, Josef Sulc, Michael Tieu, Amy Torkelson, Herman Tung, Xinxin Wang, Fangming Xie, Anna Marie Yanny, Renee Zhang, Seth A. Ament, M. Margarita Behrens, Hector Corrada Bravo, Jerold Chun, Alexander Dobin, Jesse Gillis, Ronna Hertzano, Patrick R. Hof, Thomas Höllt, Gregory D. Horwitz, C. Dirk Keene, Peter V. Kharchenko, Andrew L. Ko, Boudewijn P. Lelieveldt, Chongyuan Luo, Eran A. Mukamel, António Pinto-Duarte, Sebastian Preiss, Aviv Regev, Bing Ren, Richard H. Scheuermann, Kimberly Smith, William J. Spain, Owen R. White, Christof Koch, Michael Hawrylycz, Bosiljka Tasic, Evan Z. Macosko, Steven A. McCarroll, Jonathan T. Ting, Hongkui Zeng, Kun Zhang, Guoping Feng, Joseph R. Ecker, Sten Linnarsson, and Ed S. Lein
- Subjects
Multidisciplinary - Published
- 2022
16. Contributed Session II: Motion/direction-sensitive thalamic neurons project extensively to the middle layers of primary visual cortex
- Author
-
Jun Zhuang, Yun Wang, Naveen D. Ouellette, Emily Turschak, Rylan S. Larsen, Kevin T. Takasaki, Tanya L. Daigle, Bosiljka Tasic, Jack Waters, Hongkui Zeng, and R. Clay Reid
- Subjects
Ophthalmology ,Sensory Systems - Published
- 2022
17. Morphological diversity of single neurons in molecularly defined cell types
- Author
-
Anan Li, Elise Shen, Lijuan Liu, Wan Wan, Hui Gong, Yanjun Duan, Rachel A. Dalley, Sujun Zhao, Luke Esposito, Zhixi Yun, Shaoqun Zeng, An Liu, Susan M. Sunkin, Zhi Zhou, Tanya L. Daigle, Jintao Pan, Liya Ding, Yaoyao Li, Chris Hill, Yimin Wang, Yefeng Zheng, Qingming Luo, Phil Lesnar, Karla E. Hirokawa, Zijun Zhao, Christof Koch, Qi Li, Ping He, Donghuan Lu, Staci A. Sorensen, Longfei Li, Zhongze Gu, Xiangning Li, Zhangcan Ding, Lei Qu, Jia Yuan, Hsien-Chi Kuo, Aaron Feiner, Stephanie Mok, Julie A. Harris, Jing Yuan, Yang Yu, Qiang Ouyang, Z. Josh Huang, X. William Yang, Guodong Hong, Thuc Nghi Nguyen, Rachael Larsen, Michael Hawrylycz, Wenjie Xu, Peng Wang, Chao Chen, Wei Xiong, Hongkui Zeng, Mengya Chen, Zongcai Ruan, Feng Xiong, Shichen Zhang, Lydia Ng, Min Ye, Wayne Wakeman, Peng Xie, Yaping Wang, Quanxin Wang, Yun Wang, Sara Kebede, Bosiljka Tasic, Lulu Yin, Yuanyuan Song, Tao Wang, Lei Huang, Wei Xie, Zizhen Yao, Matthew B. Veldman, Yuanyuan Li, Xiuli Kuang, Shengdian Jiang, and Hanchuan Peng
- Subjects
Cell type ,Neurogenesis ,Neocortex ,Biology ,Neural circuits ,Article ,Atlases as Topic ,Cellular neuroscience ,Biological neural network ,medicine ,Feature (machine learning) ,Humans ,RNA-Seq ,Axon ,Projection (set theory) ,Cell Shape ,Neurons ,Multidisciplinary ,Brain ,Gene Expression Regulation, Developmental ,Reproducibility of Results ,Claustrum ,medicine.anatomical_structure ,Evolutionary biology ,Single-Cell Analysis ,Neuroglia ,Function (biology) ,Biomarkers - Abstract
Dendritic and axonal morphology reflects the input and output of neurons and is a defining feature of neuronal types1,2, yet our knowledge of its diversity remains limited. Here, to systematically examine complete single-neuron morphologies on a brain-wide scale, we established a pipeline encompassing sparse labelling, whole-brain imaging, reconstruction, registration and analysis. We fully reconstructed 1,741 neurons from cortex, claustrum, thalamus, striatum and other brain regions in mice. We identified 11 major projection neuron types with distinct morphological features and corresponding transcriptomic identities. Extensive projectional diversity was found within each of these major types, on the basis of which some types were clustered into more refined subtypes. This diversity follows a set of generalizable principles that govern long-range axonal projections at different levels, including molecular correspondence, divergent or convergent projection, axon termination pattern, regional specificity, topography, and individual cell variability. Although clear concordance with transcriptomic profiles is evident at the level of major projection type, fine-grained morphological diversity often does not readily correlate with transcriptomic subtypes derived from unsupervised clustering, highlighting the need for single-cell cross-modality studies. Overall, our study demonstrates the crucial need for quantitative description of complete single-cell anatomy in cell-type classification, as single-cell morphological diversity reveals a plethora of ways in which different cell types and their individual members may contribute to the configuration and function of their respective circuits., Sparse labelling and whole-brain imaging are used to reconstruct and classify brain-wide complete morphologies of 1,741 individual neurons in the mouse brain, revealing a dependence on both brain region and transcriptomic profile.
- Published
- 2020
18. Alternating sources of perisomatic inhibition during behavior
- Author
-
Jordane Dimidschstein, Attila Losonczy, Olivia Fong, John C. Bowler, Hongkui Zeng, Brian Lee, Gergely G. Szabo, Ivan Soltesz, Jordan S. Farrell, Ernie Hwaun, Jim Berg, Bosiljka Tasic, Peter M. Klein, Zizhen Yao, Barna Dudok, Fraser T. Sparks, Tanya L. Daigle, Satoshi Terada, and Gord Fishell
- Subjects
0301 basic medicine ,Male ,Interneuron ,Hippocampus ,Mice, Transgenic ,Hippocampal formation ,Inhibitory postsynaptic potential ,Synaptic Transmission ,03 medical and health sciences ,0302 clinical medicine ,Basket cell ,Interneurons ,medicine ,Animals ,CA1 Region, Hippocampal ,biology ,Chemistry ,General Neuroscience ,Pyramidal Cells ,digestive, oral, and skin physiology ,Mice, Inbred C57BL ,030104 developmental biology ,medicine.anatomical_structure ,Parvalbumins ,nervous system ,biology.protein ,GABAergic ,Female ,Pyramidal cell ,Cholecystokinin ,Neuroscience ,hormones, hormone substitutes, and hormone antagonists ,030217 neurology & neurosurgery ,Parvalbumin - Abstract
Summary Interneurons expressing cholecystokinin (CCK) and parvalbumin (PV) constitute two key GABAergic controllers of hippocampal pyramidal cell output. Although the temporally precise and millisecond-scale inhibitory regulation of neuronal ensembles delivered by PV interneurons is well established, the in vivo recruitment patterns of CCK-expressing basket cell (BC) populations has remained unknown. We show in the CA1 of the mouse hippocampus that the activity of CCK BCs inversely scales with both PV and pyramidal cell activity at the behaviorally relevant timescales of seconds. Intervention experiments indicated that the inverse coupling of CCK and PV GABAergic systems arises through a mechanism involving powerful inhibitory control of CCK BCs by PV cells. The tightly coupled complementarity of two key microcircuit regulatory modules demonstrates a novel form of brain-state-specific segregation of inhibition during spontaneous behavior.
- Published
- 2020
19. Evolution of cellular diversity in primary motor cortex of human, marmoset monkey, and mouse
- Author
-
Trygve E. Bakken, Nikolas L. Jorstad, Qiwen Hu, Blue B. Lake, Wei Tian, Brian E. Kalmbach, Megan Crow, Rebecca D. Hodge, Fenna M. Krienen, Staci A. Sorensen, Jeroen Eggermont, Zizhen Yao, Brian D. Aevermann, Andrew I. Aldridge, Anna Bartlett, Darren Bertagnolli, Tamara Casper, Rosa G. Castanon, Kirsten Crichton, Tanya L. Daigle, Rachel Dalley, Nick Dee, Nikolai Dembrow, Dinh Diep, Song-Lin Ding, Weixiu Dong, Rongxin Fang, Stephan Fischer, Melissa Goldman, Jeff Goldy, Lucas T. Graybuck, Brian R. Herb, Xiaomeng Hou, Jayaram Kancherla, Matthew Kroll, Kanan Lathia, Baldur van Lew, Yang Eric Li, Christine S. Liu, Hanqing Liu, Jacinta D. Lucero, Anup Mahurkar, Delissa McMillen, Jeremy A. Miller, Marmar Moussa, Joseph R. Nery, Philip R. Nicovich, Joshua Orvis, Julia K. Osteen, Scott Owen, Carter R. Palmer, Thanh Pham, Nongluk Plongthongkum, Olivier Poirion, Nora M. Reed, Christine Rimorin, Angeline Rivkin, William J. Romanow, Adriana E. Sedeño-Cortés, Kimberly Siletti, Saroja Somasundaram, Josef Sulc, Michael Tieu, Amy Torkelson, Herman Tung, Xinxin Wang, Fangming Xie, Anna Marie Yanny, Renee Zhang, Seth A. Ament, M. Margarita Behrens, Hector Corrada Bravo, Jerold Chun, Alexander Dobin, Jesse Gillis, Ronna Hertzano, Patrick R. Hof, Thomas Höllt, Gregory D. Horwitz, C. Dirk Keene, Peter V. Kharchenko, Andrew L. Ko, Boudewijn P. Lelieveldt, Chongyuan Luo, Eran A. Mukamel, Sebastian Preissl, Aviv Regev, Bing Ren, Richard H. Scheuermann, Kimberly Smith, William J. Spain, Owen R. White, Christof Koch, Michael Hawrylycz, Bosiljka Tasic, Evan Z. Macosko, Steven A. McCarroll, Jonathan T. Ting, Hongkui Zeng, Kun Zhang, Guoping Feng, Joseph R. Ecker, Sten Linnarsson, and Ed S. Lein
- Subjects
Transcriptome ,Cell type ,biology ,Evolutionary biology ,biology.animal ,DNA methylation ,Marmoset ,Epigenome ,Gene ,Chromatin ,Epigenomics - Abstract
The primary motor cortex (M1) is essential for voluntary fine motor control and is functionally conserved across mammals. Using high-throughput transcriptomic and epigenomic profiling of over 450,000 single nuclei in human, marmoset monkey, and mouse, we demonstrate a broadly conserved cellular makeup of this region, whose similarity mirrors evolutionary distance and is consistent between the transcriptome and epigenome. The core conserved molecular identity of neuronal and non-neuronal types allowed the generation of a cross-species consensus cell type classification and inference of conserved cell type properties across species. Despite overall conservation, many species specializations were apparent, including differences in cell type proportions, gene expression, DNA methylation, and chromatin state. Few cell type marker genes were conserved across species, providing a short list of candidate genes and regulatory mechanisms responsible for conserved features of homologous cell types, such as the GABAergic chandelier cells. This consensus transcriptomic classification allowed the Patch-seq identification of layer 5 (L5) corticospinal Betz cells in non-human primate and human and characterization of their highly specialized physiology and anatomy. These findings highlight the robust molecular underpinnings of cell type diversity in M1 across mammals and point to the genes and regulatory pathways responsible for the functional identity of cell types and their species-specific adaptations.
- Published
- 2020
- Full Text
- View/download PDF
20. Toward an integrated classification of neuronal cell types: morphoelectric and transcriptomic characterization of individual GABAergic cortical neurons
- Author
-
David Feng, Jessica Trinh, Tamara Casper, Lisa Kim, Rohan Gala, Clare Gamlin, Matthew Kroll, Uygar Sümbül, Lauren Alfiler, Thomas Braun, Jasmine Bomben, Bosiljka Tasic, Colin Farrell, Hongkui Zeng, Lydia Potekhina, Tsega Desta, Kiet Ngo, Lydia Ng, Alice Mukora, Fahimeh Baftizadeh, Aaron Szafer, Rachel A. Dalley, Shea Ransford, Changkyu Lee, Nick Dee, Brian Lee, Kirsten Crichton, Luke Esposito, Miranda Robertson, Josef Sulc, Alex M. Henry, Darren Bertagnolli, Tom Egdorf, Nadezhda Dotson, Zhi Zhou, Jim Berg, Philip R. Nicovich, Rusty Mann, Madie Hupp, Daniel Park, Delissa McMillen, Samuel Dingman Lee, Agata Budzillo, Eliza Barkan, Olivia Fong, Thanh Pham, Jeff Goldy, Ed S. Lein, Rebecca de Frates, Kimberly A. Smith, Amy Torkelson, Tim Jarsky, Michelle Maxwell, Michael Tieu, Susan M. Sunkin, Michael Hawrylycz, Lucas T. Graybuck, Herman Tung, David Reid, DiJon Hill, Alexandra Glandon, Kara Ronellenfitch, Aaron Oldre, Amanda Gary, Nathan W. Gouwens, Christof Koch, Alice Pom, Wayne Wakeman, Sara Kebede, Matthew Mallory, Tae Kyung Kim, Tanya L. Daigle, Kris Bickley, Anton Arkhipov, Osnat Penn, Staci A. Sorensen, Rachel Enstrom, Hanchuan Peng, Ramkumar Rajanbabu, Jonathan T. Ting, Zizhen Yao, Lauren Ellingwood, Medea McGraw, Gabe J. Murphy, Katherine Baker, Krissy Brouner, Hong Gu, David Sandman, Katelyn Ward, Kyla Berry, Katherine E. Link, Lindsay Ng, Christine Rimorin, Kristen Hadley, Augustin Ruiz, Grace Williams, and Melissa Gorham
- Subjects
Transcriptome ,Electrophysiology ,Cell type ,medicine.anatomical_structure ,Visual cortex ,Interneuron ,nervous system ,genetic structures ,medicine ,GABAergic ,Cortical neurons ,Biology ,Neuroscience - Abstract
Neurons are frequently classified into distinct groups or cell types on the basis of structural, physiological, or genetic attributes. To better constrain the definition of neuronal cell types, we characterized the transcriptomes and intrinsic physiological properties of over 3,700 GABAergic mouse visual cortical neurons and reconstructed the local morphologies of 350 of those neurons. We found that most transcriptomic types (t-types) occupy specific laminar positions within mouse visual cortex, and many of those t-types exhibit consistent electrophysiological and morphological features. We observed that these properties could vary continuously between t-types, which limited the ability to predict specific t-types from other data modalities. Despite that, the data support the presence of at least 20 interneuron met-types that have congruent morphological, electrophysiological, and transcriptomic properties.HighlightsPatch-seq data obtained from >3,700 GABAergic cortical interneuronsComprehensive characterization of morpho-electric features of transcriptomic types20 interneuron met-types that have congruent properties across data modalitiesDifferent Sst met-types preferentially innervate different cortical layers
- Published
- 2020
- Full Text
- View/download PDF
21. Toward an Integrated Classification of Cell Types: Morphoelectric and Transcriptomic Characterization of Individual GABAergic Cortical Neurons
- Author
-
Kimberly A. Smith, Matthew Kroll, Sara Kebede, Susan M. Sunkin, David Reid, Nadezhda Dotson, Rusty Mann, DiJon Hill, Kara Ronellenfitch, Shea Ransford, Hongkui Zeng, David Feng, Jasmine Bomben, Bosiljka Tasic, Rachel Enstrom, Jessica Trinh, Matthew Mallory, Aaron Szafer, Rachel A. Dalley, Aaron Oldre, Amanda Gary, Eliza Barkan, Nick Dee, Lydia Ng, Tae Kyung Kim, Ed S. Lein, Colin Farrell, Tamara Casper, Tom Egdorf, Kirsten Crichton, Josef Sulc, Fahimeh Baftizadeh, Katelyn Ward, Kirsten Hadley, Alex M. Henry, Alice Pom, Brian Lee, Uygar Sümbül, Lisa Kim, Tim Jarsky, Madie Happ, Wayne Wakeman, Lauren Ellingwood, Luke Esposito, Daniel Park, Tanya L. Daigle, Darren Bertagnolli, Lucas T. Graybuck, Olivia Fong, Philip R. Nicovich, Gabe J. Murphy, Michelle Maxwell, Lindsay Ng, Rebeeca de Frates, Rohan Gala, Alice Mukora, Delissa McMillen, Miranda Robertson, Thanh Pham, Samuel Dingman Lee, Kris Bickley, Anton Arkhipov, Osnat Penn, Staci A. Sorensen, Alexandra Glandon, Zizhen Yao, Amy Torkelson, Jonathan T. Ting, Lauren Alfiler, Ramkumar Rajanbabu, Kiet Ngo, Kirssy Brouner, David Sandman, Michael Tieu, Michael Hawrylycz, Nathan W. Gouwens, Hanchuan Peng, Zhi Zhou, Jeff Goldy, Hong Gu, Herman Tung, Medea McGraw, Lyida Potekhina, Katherine Baker, Tsega Desta, Christof Koch, Changkyu Lee, Melissa Gorham, Clare Gamlin, Augustin Ruiz, Grace Williams, Jim Berg, Kyla Berry, Katherine E. Link, Agata Budzillo, Christine Rimorin, and Thomas Braun
- Subjects
Cell type ,genetic structures ,Interneuron ,Cortical neurons ,Biology ,Transcriptome ,Electrophysiology ,medicine.anatomical_structure ,Visual cortex ,nervous system ,medicine ,biology.protein ,GABAergic ,Neuroscience ,Parvalbumin - Abstract
Neurons are frequently classified into distinct groups or cell types on the basis of structural, physiological, or genetic attributes. To better constrain the definition of neuronal cell types, we characterized the transcriptomes and intrinsic physiological properties of over 3,700 GABAergic mouse visual cortical neurons and reconstructed the local morphologies of 350 of those neurons. We found that most transcriptomic types (t-types) occupy specific laminar positions within mouse visual cortex, and many of those t-types exhibit consistent electrophysiological and morphological features. We observed that these properties could vary continuously between t- types, which limited the ability to predict specific t-types from other data modalities. Despite that, the data support the presence of at least 20 interneuron met-types that have congruent morphological, electrophysiological, and transcriptomic properties.
- Published
- 2020
22. The spatial structure of feedforward information in mouse primary visual cortex
- Author
-
Tanya L. Daigle, Bosiljka Tasic, Jun Zhuang, Hongkui Zeng, Rylan S. Larsen, Jack Waters, R. Clay Reid, Naveen D. Ouellette, and Kevin T. Takasaki
- Subjects
medicine.anatomical_structure ,Visual cortex ,nervous system ,Receptive field ,Spatial structure ,medicine ,Motion direction ,Feed forward ,Spatial frequency ,Multiple modalities ,Biology ,Nucleus ,Neuroscience - Abstract
Location-sensitive and motion-sensitive units are the two major functional types of feedforward projections from lateral genicular nucleus (LGN) to primary visual cortex (V1) in mouse. The distribution of these inputs in cortical depth remains under debate. By measuring the calcium activities of LGN axons in V1 of awake mice, we systematically mapped their functional and structural properties. Although both types distributed evenly across cortical depth, we found that they differ significantly across multiple modalities. Compared to the location-sensitive axons, which possessed confined spatial receptive fields, the motion-sensitive axons lacked spatial receptive fields, preferred lower temporal, higher spatial frequencies and had wider horizontal bouton spread. Furthermore, the motion-sensitive axons showed a strong depth-dependent motion direction bias while the location-sensitive axons showed a depth-independent OFF dominance. Overall, our results suggest a new model of receptive biases and laminar structure of thalamic inputs to V1.
- Published
- 2019
23. Signature morpho-electric, transcriptomic, and dendritic properties of human layer 5 neocortical pyramidal neurons
- Author
-
Brian Lee, Rachel A. Dalley, Andrew L. Ko, Nikolas L. Jorstad, Anoop P. Patel, Jeffrey G. Ojemann, Lucas T. Graybuck, Brian E. Kalmbach, Rebecca D. Hodge, Kimberly A. Smith, Tanya L. Daigle, Anna Marie Yanny, Hongkui Zeng, Philip R. Nicovich, Cristina Radaelli, Richard G. Ellenbogen, Daniel L. Silbergeld, Matt Mallory, Nick Dee, Scott F. Owen, Christof Koch, Ed S. Lein, Rebecca de Frates, Staci A. Sorensen, C. Dirk Keene, Medea McGraw, Trygve E. Bakken, Charles Cobbs, Bosiljka Tasic, Jonathan T. Ting, and Ryder P. Gwinn
- Subjects
Adult ,Male ,Cell type ,Patch-Clamp Techniques ,Action Potentials ,Mice, Transgenic ,Neocortex ,Dendrite ,Biology ,Article ,Transcriptome ,Mice ,Organ Culture Techniques ,Morphogenesis ,medicine ,Animals ,Humans ,Dendritic spike ,Pyramidal Cells ,General Neuroscience ,Functional specialization ,Dendrites ,Middle Aged ,Phenotype ,Mice, Inbred C57BL ,medicine.anatomical_structure ,nervous system ,Female ,Macaca nemestrina ,Neuroscience ,Function (biology) - Abstract
In the neocortex, subcerebral axonal projections originate largely from layer 5 (L5) extratelencephalic-projecting (ET) neurons. The unique morpho-electric properties of these neurons have been mainly described in rodents, where retrograde tracers or transgenic lines can label them. Similar labeling strategies are infeasible in the human neocortex, rendering the translational relevance of findings in rodents unclear. We leveraged the recent discovery of a transcriptomically defined L5 ET neuron type to study the properties of human L5 ET neurons in neocortical brain slices derived from neurosurgeries. Patch-seq recordings, where transcriptome, physiology, and morphology were assayed from the same cell, revealed many conserved morpho-electric properties of human and rodent L5 ET neurons. Divergent properties were often subtler than differences between L5 cell types within these two species. These data suggest a conserved function of L5 ET neurons in the neocortical hierarchy but also highlight phenotypic divergence possibly related to functional specialization of human neocortex.
- Published
- 2021
24. Sirtuin 5 protects mitochondria from fragmentation and degradation during starvation
- Author
-
Hala Guedouari, Luca Scorrano, Tanya L. Daigle, and Etienne Hebert-Chatelain
- Subjects
Dynamins ,0301 basic medicine ,SIRT3 ,Mitochondrial Degradation ,Biology ,Mitochondrion ,Mitochondrial Dynamics ,Mitochondrial apoptosis-induced channel ,Mice ,03 medical and health sciences ,DNM1L ,0302 clinical medicine ,Stress, Physiological ,Sirtuin 5 ,Autophagy ,Animals ,Sirtuins ,Molecular Biology ,Mitochondrial degradation ,Cell Line, Transformed ,2. Zero hunger ,Mitochondrial fragmentation ,Mitophagy ,Cell Biology ,Fibroblasts ,Protective Factors ,Culture Media ,Mitochondria ,Cell biology ,Glucose ,030104 developmental biology ,Biochemistry ,DNAJA3 ,Mitochondrial fission ,ATP–ADP translocase ,Gene Deletion ,030217 neurology & neurosurgery - Abstract
During starvation, intra-mitochondrial sirtuins, NAD+ sensitive deacylating enzymes that modulate metabolic homeostasis and survival, directly adjust mitochondrial function to nutrient availability; concomitantly, mitochondria elongate to escape autophagic degradation. However, whether sirtuins also impinge on mitochondrial dynamics is still uncharacterized. Here we show that the mitochondrial Sirtuin 5 (Sirt5) is essential for starvation induced mitochondrial elongation. Deletion of Sirt5 in mouse embryonic fibroblasts increased levels of mitochondrial dynamics of 51kDa protein and mitochondrial fission protein 1, leading to mitochondrial accumulation of the pro-fission dynamin related protein 1 and to mitochondrial fragmentation. During starvation, Sirt5 deletion blunted mitochondrial elongation, resulting in increased mitophagy. Our results indicate that starvation induced mitochondrial elongation and evasion from autophagic degradation requires the energy sensor Sirt5.
- Published
- 2017
25. Visual Cortex Gains Independence from Peripheral Drive before Eye Opening
- Author
-
Alexandra Gribizis, Hongkui Zeng, Michael C. Crair, Tanya L. Daigle, James B. Ackman, Daeyeol Lee, and Xinxin Ge
- Subjects
0301 basic medicine ,Male ,Superior Colliculi ,genetic structures ,Neurogenesis ,Thalamus ,Sensory system ,Biology ,Lateral geniculate nucleus ,Article ,Retina ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Cortex (anatomy) ,medicine ,Premovement neuronal activity ,Animals ,Visual Pathways ,Visual Cortex ,Neurons ,General Neuroscience ,Superior colliculus ,Retinal waves ,Mice, Inbred C57BL ,030104 developmental biology ,medicine.anatomical_structure ,Visual cortex ,Female ,Neuroscience ,030217 neurology & neurosurgery - Abstract
Summary Visual spatial perception in the mammalian brain occurs through two parallel pathways: one reaches the primary visual cortex (V1) through the thalamus and another the superior colliculus (SC) via direct projections from the retina. The origin, development, and relative function of these two evolutionarily distinct pathways remain obscure. We examined the early functional development of both pathways by simultaneously imaging pre- and post-synaptic spontaneous neuronal activity. We observed that the quality of retinal activity transfer to the thalamus and superior colliculus does not change across the first two postnatal weeks. However, beginning in the second postnatal week, retinal activity does not drive V1 as strongly as earlier wave activity, suggesting that intrinsic cortical activity competes with signals from the sensory periphery as the cortex matures. Together, these findings bring new insight into the function of the SC and V1 and the role of peripheral activity in driving both circuits across development.
- Published
- 2019
26. RecV recombinase system for spatiotemporally controlled light-inducible genomic modifications
- Author
-
Yun Wang, Bosiljka Tasic, Shenqin Yao, Viviana Gradinaru, Mark J. Schnitzer, Pooja Balaram, Andrew Curtright, Anat Kahan, Thomas Zhou, Radosław Chrapkiewicz, Peng Yuan, Soumya Chatterjee, Ajay Dhaka, Ali Cetin, Hui Gong, Hongkui Zeng, Shaoqun Zeng, Xiuli Kuang, Tanya L. Daigle, Qingming Luo, Ben Ouellette, and Marty Mortrud
- Subjects
0303 health sciences ,Cell ,Computational biology ,Biology ,Inhibitory postsynaptic potential ,Viral vector ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,medicine.anatomical_structure ,chemistry ,Recombinase ,medicine ,Biological neural network ,Soma ,030217 neurology & neurosurgery ,Function (biology) ,DNA ,030304 developmental biology - Abstract
Brain circuits are composed of vast numbers of intricately interconnected neurons with diverse molecular, anatomical and physiological properties. To allow highly specific “user-defined” targeting of individual neurons for structural and functional studies, we modified three site-specific DNA recombinases, Cre, Dre and Flp, by combining them with a fungal light-inducible protein, Vivid, to create light-inducible recombinases (named RecV). We generated viral vectors to express these light-inducible recombinases and demonstrated that they can induce genomic modifications in dense or sparse populations of neurons in superficial as well as deep brain areas of live mouse brains by one-photon or two-photon light induction. These light-inducible recombinases can produce highly targeted, sparse and strong labeling of individual neurons in multiple loci and species. They can be used in combination with other genetic strategies to achieve specific intersectional targeting of mouse cortical layer 5 or inhibitory somatostatin neurons. In mouse cortex sparse light-induced recombination allows whole-brain morphological reconstructions to identify axonal projection specificity. Furthermore these enzymes allow single cell targeted genetic modifications via soma restricted two-photon light stimulation in individual cortical neurons and can be used in combination with functional optical indicators with minimal interference. In summary, RecVs enable spatiotemporally-precise, targeted optogenomic modifications that could greatly facilitate detailed analysis of neural circuits at the single cell level by linking genetic identity, morphology, connectivity and function.
- Published
- 2019
- Full Text
- View/download PDF
27. Complete Whole-Brain Single Neuron Reconstruction Reveals Morphological Diversity in Molecularly Defined Claustral and Cortical Neuron Types
- Author
-
Z. Josh Huang, Matthew B. Veldman, Yang Yu, Sara Kebede, Philip Lesnar, Thuc Nghi Nguyen, Anan Li, Zhi Zhou, Chris Hill, Susan M. Sunkin, Hanchuan Peng, Shaoqun Zeng, Yaoyao Li, Qingming Luo, Li Xiangning, Lei Qu, Karla E. Hirokawa, Elise Shen, Lijuan Liu, Yimin Wang, X. William Yang, Michael Hawrylycz, Lydia Ng, Hui Gong, Peng Xie, Sujun Zhao, Xiuli Kuang, Tanya L. Daigle, Aaron Feiner, Zizhen Yao, Christof Koch, Shengdian Jiang, Stephanie Mok, Jing Yuan, Hongkui Zeng, Lulu Ying, Rachael Larsen, Staci A. Sorensen, Julie A. Harris, Luke Esposito, Yun Wang, Bosiljka Tasic, Yuanyuan Song, Quanxin Wang, and Wayne Wakeman
- Subjects
Fluorescent labelling ,medicine.anatomical_structure ,Feature (computer vision) ,Cortical neuron ,Cortex (anatomy) ,medicine ,Neuron ,Biology ,Fluorescent imaging ,Neuroscience ,Claustrum - Abstract
Dendritic and axonal morphology is a defining feature of neuronal types and their connectivity. Yet our knowledge concerning the diversity of neuronal morphology is extremely limited. To systematically obtain single neuron full morphology on a brain-wide scale in mice, we established a pipeline that encompasses five major components: sparse labeling, whole-brain imaging, reconstruction, registration, and classification. We achieved sparse, robust and consistent fluorescent labeling by combining transgenic or viral Cre delivery with novel transgenic reporter lines, and generated whole-brain fluorescent imaging datasets containing tens of thousands of reconstructable neurons. We developed software tools for large-volume image data processing and computer-assisted morphological reconstruction. For a proof-of-principle, we reconstructed the full morphologies of 96 neurons from the claustrum and cortex that belong to a single transcriptomically-defined subclass, and classified them into multiple morphological types, suggesting that they work in a targeted and coordinated manner to process cortical information over a large region.
- Published
- 2019
28. Activation of neuromodulatory axon projections in primary visual cortex during periods of locomotion and pupil dilation
- Author
-
Tanya L. Daigle, Jack Waters, Emily Turschak, Rylan S. Larsen, Jun Zhuang, and Hongkui Zeng
- Subjects
genetic structures ,Biology ,eye diseases ,Arousal ,Norepinephrine ,medicine.anatomical_structure ,Visual cortex ,nervous system ,Cortex (anatomy) ,medicine ,Pupillary response ,Cholinergic ,sense organs ,Axon ,Neuroscience ,Acetylcholine ,medicine.drug - Abstract
Neuromodulators such as acetylcholine, noradrenaline (norepinephrine), and serotonin are released into the cortex by axons ascending from subcortical nuclei. These neuromodulators have been hypothesized to influence cortical function during behavioral periods such as arousal, locomotion, exploration, and attention. To determine when these neuromodulatory projections were active, we expressed the genetically-encoded calcium sensor GCaMP6 in neuromodulatory axons which project to the mouse primary visual cortex and performed two-photon microscopy to monitor their activity in vivo. We observed that the fluorescence of both cholinergic and noradrenergic axons increased during periods of pupil dilation, with the fluorescence of the axons rising less than one second before eye pupil dilation. We also observed increases in cholinergic and noradrenergic axon fluorescence periods of locomotion, which was accompanied by pupil dilation and nasal (forward) movement of both pupils. Locomotion was preceded by a rise in axonal fluorescence with a timing and amplitude that matched the subsequent pupil dilation, but axon fluorescence was more sustained than expected from the pupil dilation, suggesting that there is an additional physiological factor that affects cholinergic and noradrenergic axon activity in primary visual cortex during locomotion.
- Published
- 2018
29. Integrated Morphoelectric and Transcriptomic Classification of Cortical GABAergic Cells
- Author
-
Kris Bickley, Anton Arkhipov, Osnat Penn, Hanchuan Peng, Shea Ransford, Sara Kebede, Kara Ronellenfitch, Matthew Mallory, Krissy Brouner, Madie Hupp, Lydia Ng, Daniel Park, Staci A. Sorensen, Alice Pom, Susan M. Sunkin, Tanya L. Daigle, Fahimeh Baftizadeh, Wayne Wakeman, Aaron Oldre, Amanda Gary, Herman Tung, Brian Lee, Ed S. Lein, Medea McGraw, Rachel A. Dalley, Bosiljka Tasic, Hong Gu, Miranda Robertson, Katherine Baker, Lindsay Ng, David Sandman, Jasmine Bomben, Uygar Sümbül, Tae Kyung Kim, David Reid, Eliza Barkan, Luke Esposito, Kirsten Crichton, DiJon Hill, Zoran Popović, Josef Sulc, Nathan W. Gouwens, Ramkumar Rajanbabu, Lydia Potekhina, Thomas Braun, Alexandra Glandon, Tim Jarsky, Darren Bertagnolli, Tom Egdorf, Olivia Fong, Alice Mukora, Rebecca de Frates, Lauren Ellingwood, Jonathan T. Ting, Gabe J. Murphy, Katelyn Ward, Delissa McMillen, Samuel Dingman Lee, Melissa Gorham, Michelle Maxwell, Clare Gamlin, Zhi Zhou, Jeff Goldy, Rachel Enstrom, Kyla Berry, Colin Farrell, Katherine E. Link, Christine Rimorin, Zizhen Yao, Hongkui Zeng, Kristen Hadley, Augustin Ruiz, Grace Williams, Amy Torkelson, Kimberly A. Smith, Lisa Kim, Aaron Szafer, Nick Dee, Alex M. Henry, Rohan Gala, David Feng, Jessica Trinh, Tamara Casper, Matthew Kroll, Christof Koch, Michael Tieu, Michael Hawrylycz, Lauren Alfiler, Kiet Ngo, Philip R. Nicovich, Thanh Pham, Nadezhda Dotson, Rusty Mann, Tsega Desta, Lucas T. Graybuck, Changkyu Lee, Jim Berg, and Agata Budzillo
- Subjects
0303 health sciences ,Cell type ,biology ,Interneuron ,General Biochemistry, Genetics and Molecular Biology ,Transcriptome ,03 medical and health sciences ,Electrophysiology ,0302 clinical medicine ,medicine.anatomical_structure ,Visual cortex ,medicine ,biology.protein ,GABAergic ,Axon ,Neuroscience ,030217 neurology & neurosurgery ,Parvalbumin ,030304 developmental biology - Abstract
Neurons are frequently classified into distinct types on the basis of structural, physiological, or genetic attributes. To better constrain the definition of neuronal cell types, we characterized the transcriptomes and intrinsic physiological properties of over 4,200 mouse visual cortical GABAergic interneurons and reconstructed the local morphologies of 517 of those neurons. We find that most transcriptomic types (t-types) occupy specific laminar positions within visual cortex, and, for most types, the cells mapping to a t-type exhibit consistent electrophysiological and morphological properties. These properties display both discrete and continuous variation among t-types. Through multimodal integrated analysis, we define 28 met-types that have congruent morphological, electrophysiological, and transcriptomic properties and robust mutual predictability. We identify layer-specific axon innervation pattern as a defining feature distinguishing different met-types. These met-types represent a unified definition of cortical GABAergic interneuron types, providing a systematic framework to capture existing knowledge and bridge future analyses across different modalities.
- Published
- 2020
30. Brainwide Genetic Sparse Cell Labeling to Illuminate the Morphology of Neurons and Glia with Cre-Dependent MORF Mice
- Author
-
Hong-Wei Dong, Ivan A. Lopez, Elizabeth Zuniga-Sanchez, Chang Sin Park, Matthew B. Veldman, Tanya L. Daigle, Muye Zhu, S. Lawrence Zipursky, Charles M. Eyermann, Arlene A. Hirano, Peter Langfelder, X. William Yang, Jason Y. Zhang, Nicholas C. Brecha, Nicholas N. Foster, and Hongkui Zeng
- Subjects
0301 basic medicine ,microglia ,Morphology (biology) ,Retinal Horizontal Cells ,Cell morphology ,Transgenic ,Mice ,0302 clinical medicine ,morphology ,Psychology ,Frameshift Mutation ,Neurons ,Microglia ,General Neuroscience ,Brain ,imaging ,Cre ,MORF ,medicine.anatomical_structure ,Neurological ,Cognitive Sciences ,Astrocyte ,reconstruction ,sparse labeling ,1.1 Normal biological development and functioning ,Green Fluorescent Proteins ,Mice, Transgenic ,Biology ,Article ,Cell labeling ,Frameshift mutation ,03 medical and health sciences ,astrocyte ,Underpinning research ,Genetics ,medicine ,Animals ,Neurology & Neurosurgery ,Integrases ,Neurosciences ,neuron ,spaghetti monster ,Brain Disorders ,030104 developmental biology ,nervous system ,Astrocytes ,Neuron ,Neuroscience ,030217 neurology & neurosurgery ,Function (biology) ,Microsatellite Repeats - Abstract
Cajal recognized that the elaborate shape of neurons is fundamental to their function in the brain. However, there are no simple and generalizable genetic methods to study neuronal or glial cell morphology in the mammalian brain. Here, we describe four mouse lines conferring Cre-dependent sparse cell labeling based on mononucleotide repeat frameshift (MORF) as a stochastic translational switch. Notably, the optimized MORF3 mice, with a membrane-bound multivalent immunoreporter, confer Cre-dependent sparse and bright labeling of thousands of neurons, astrocytes, or microglia in each brain, revealing their intricate morphologies. MORF3 mice are compatible with imaging in tissue-cleared thick brain sections and with immuno-EM. An analysis of 151 MORF3-labeled developing retinal horizontal cells reveals novel morphological cellclusters and axonal maturation patterns. Our study demonstrates a conceptually novel, simple, generalizable, and scalable mouse genetic solution to sparsely label and illuminate the morphology of genetically defined neurons and glia in the mammalian brain.
- Published
- 2020
31. Classification of electrophysiological and morphological types in mouse visual cortex
- Author
-
Tom Egdorf, Rebecca de Frates, Emma Garren, Sara Kebede, Peter Chong, John W. Phillips, Nivretta Thatra, Samuel R Josephsen, Philip R. Nicovich, Tim Jarsky, Xiaoxiao Liu, Susan M. Sunkin, Brian Lee, Keith B. Godfrey, Matthew Kroll, Nicole Blesie, Bosiljka Tasic, Amy Bernard, Lisa Kim, Costas A. Anastassiou, Kristen Hadley, Staci A. Sorensen, Thuc Nghi Nguyen, Martin Schroedter, Corinne Teeter, Kirsten Crichton, Josef Sulc, Rachel A. Dalley, David Feng, Tracy Lemon, Michael Hawrylycz, Miranda Robertson, Christine Cuhaciyan, Eliza Barkan, Shiella Caldejon, Tsega Desta, Kris Bickley, Dan Castelli, Wayne Wakeman, Herman Tung, Hongkui Zeng, Grace Williams, Nadezhda Dotson, Rusty Mann, Tamara Casper, Anton Arkhipov, Daniel Park, Sheana Parry, Jed Perkins, Alyse Doperalski, Brian Long, Thomas Braun, Christof Koch, Gabe J. Murphy, Aaron Oldre, Changkyu Lee, Colin Farrell, Medea McGraw, Amanda Gary, Kiet Ngo, Melissa Gorham, Naz Taskin, Jim Berg, Samuel Dingman, Tanya L. Daigle, Agata Budzillo, Marissa Garwood, Gilberto J. Soler-Llavina, Aaron Szafer, Nick Dee, Jonathan T. Ting, Lydia Ng, Alex M. Henry, James Harrington, Julie A. Harris, Michael S. Fisher, Lindsay Ng, Caroline Habel, Nathalie Gaudreault, Krissy Brouner, David Reid, Lydia Potekhina, Rob Young, DiJon Hill, Cliff Slaughterbeck, Ed Lein, Alice Mukora, David Sandman, Stefan Mihalas, Nathan W. Gouwens, Zhi Zhou, Hanchuan Peng, and Hong Gu
- Subjects
Genetically modified mouse ,Cell type ,Cell ,Laboratory mouse ,Biology ,Electrophysiology ,chemistry.chemical_compound ,Visual cortex ,medicine.anatomical_structure ,chemistry ,Biocytin ,medicine ,Patch clamp ,Neuroscience - Abstract
Understanding the diversity of cell types in the brain has been an enduring challenge and requires detailed characterization of individual neurons in multiple dimensions. To profile morpho-electric properties of mammalian neurons systematically, we established a single cell characterization pipeline using standardized patch clamp recordings in brain slices and biocytin-based neuronal reconstructions. We built a publicly-accessible online database, the Allen Cell Types Database, to display these data sets. Intrinsic physiological and morphological properties were measured from over 1,800 neurons from the adult laboratory mouse visual cortex. Quantitative features were used to classify neurons into distinct types using unsupervised methods. We establish a taxonomy of morphologically- and electrophysiologically-defined cell types for this region of cortex with 17 e-types and 35 m-types, as well as an initial correspondence with previously-defined transcriptomic cell types using the same transgenic mouse lines.
- Published
- 2018
- Full Text
- View/download PDF
32. Parallel holographic illumination enables sub-millisecond two-photon optogenetic activation in mouse visual cortex in vivo
- Author
-
Brian Lee, Tanya L. Daigle, Emiliano Ronzitti, Eirini Papagiakoumou, I-Wen Chen, Hongkui Zeng, and Emiliani
- Subjects
Physics ,0303 health sciences ,Opsin ,Millisecond ,Holography ,Optogenetics ,law.invention ,03 medical and health sciences ,0302 clinical medicine ,Visual cortex ,medicine.anatomical_structure ,Two-photon excitation microscopy ,law ,Temporal resolution ,medicine ,Premovement neuronal activity ,Neuroscience ,030217 neurology & neurosurgery ,030304 developmental biology - Abstract
Selective control of action potential generation in individual cells from a neuronal ensemble is desirable for dissecting circuit mechanisms underlying perception and behavior. Here, by using two-photon (2P) temporally focused computer-generated holography (TF-CGH), we demonstrate optical manipulation of neuronal excitability at the supragranular layers of anesthetized mouse visual cortex. Utilizing amplified laser-pulses delivered via a localized holographic spot, our optical system achieves suprathreshold activation by exciting either of the three optogenetic actuators, ReaChR, CoChR or ChrimsonR, with brief illumination (≤ 10 ms) at moderate excitation power ((in average ≤ 0.2 mW/µm2 corresponding to ≤ 25 mW/cell). Using 2P-guided whole-cell or cell-attached recordings in positive neurons expressing respective opsin in vivo, we find that parallel illumination induces spikes of millisecond temporal resolution and sub-millisecond precision, which are preserved upon repetitive illuminations up to tens of Hz. Holographic stimulation thus enables temporally precise optogenetic activation independently of opsin’s channel kinetics. Furthermore, we demonstrate that parallel optogenetic activation can be combined with functional imaging for all-optical control of a neuronal sub-population that co-expresses the photosensitive opsin ReaChR and the calcium indicator GCaMP6s. Parallel optical control of neuronal activity with cellular resolution and millisecond temporal precision should be advantageous for investigating neuronal connections and further yielding causal links between connectivity, microcircuit dynamics, and brain functions.Significance statementRecent development of optogenetics allows probing the neuronal microcircuit with light by optically actuating genetically-encoded light-sensitive opsins expressed in the target cells. Here, we apply holographic light shaping and temporal focusing to simultaneously deliver axially-confined holographic patterns to opsin-positive cells situated in the living mouse cortex. Parallel illumination efficiently induces action potentials with high temporal resolution and precision for three opsins of different kinetics. We demonstrated all-optical experiments by extending the parallel optogenetic activation at low intensity to multiple neurons and concurrently monitoring their calcium dynamics. These results demonstrate fast and temporally precise in vivo control of a neuronal sub-population, opening new opportunities to reveal circuit mechanisms underlying brain functions.
- Published
- 2018
33. Shared and distinct transcriptomic cell types across neocortical areas
- Author
-
Daniel Hirschstein, Michael N. Economo, Allan R. Jones, Christine Rimorin, Eliza Barkan, Linda Madisen, Seana Parry, Susan M. Sunkin, Rachael Larsen, Hongkui Zeng, Tae Kyung Kim, Emma Garren, Kimberly A. Smith, Jeremy A. Miller, Osnat Penn, Olivia Fong, Sarada Viswanathan, Julie A. Harris, Bosiljka Tasic, Thuc Nghi Nguyen, Vilas Menon, Karel Svoboda, Matthew Kroll, Ed S. Lein, Peter A. Groblewski, Karla E. Hirokawa, Ali Cetin, Julie Pendergraft, Ian R. Wickersham, Tanya L. Daigle, Darren Bertagnolli, Jeff Goldy, Zizhen Yao, John W. Phillips, Michael Tieu, Loren L. Looger, Michael Hawrylycz, Aaron Szafer, Boaz P. Levi, Trygve E. Bakken, Nick Dee, Nadiya V. Shapovalova, Amy Bernard, Tamara Casper, Christof Koch, Kanan Lathia, and Lucas T. Graybuck
- Subjects
Transcriptome ,Cell type ,medicine.anatomical_structure ,Neocortex ,Visual cortex ,Cell ,medicine ,Excitatory postsynaptic potential ,Biology ,Inhibitory postsynaptic potential ,Neuroscience ,Motor cortex - Abstract
Neocortex contains a multitude of cell types segregated into layers and functionally distinct regions. To investigate the diversity of cell types across the mouse neocortex, we analyzed 12,714 cells from the primary visual cortex (VISp), and 9,035 cells from the anterior lateral motor cortex (ALM) by deep single-cell RNA-sequencing (scRNA-seq), identifying 116 transcriptomic cell types. These two regions represent distant poles of the neocortex and perform distinct functions. We define 50 inhibitory transcriptomic cell types, all of which are shared across both cortical regions. In contrast, 49 of 52 excitatory transcriptomic types were found in either VISp or ALM, with only three present in both. By combining single cell RNA-seq and retrograde labeling, we demonstrate correspondence between excitatory transcriptomic types and their region-specific long-range target specificity. This study establishes a combined transcriptomic and projectional taxonomy of cortical cell types from functionally distinct regions of the mouse cortex.
- Published
- 2017
- Full Text
- View/download PDF
34. Shared and distinct transcriptomic cell types across neocortical areas
- Author
-
Zizhen Yao, Christof Koch, Daniel Hirschstein, Aaron Szafer, Nick Dee, Sheana Parry, Michael Tieu, Michael Hawrylycz, Jeff Goldy, Susan M. Sunkin, Nadiya V. Shapovalova, Amy Bernard, Kanan Lathia, Lucas T. Graybuck, Boaz P. Levi, Trygve E. Bakken, Matthew Kroll, Sarada Viswanathan, Kimberly A. Smith, Thuc Nghi Nguyen, Olivia Fong, Tae Kyung Kim, Tanya L. Daigle, Jeremy A. Miller, Christine Rimorin, Linda Madisen, Karla E. Hirokawa, Tamara Casper, Julie A. Harris, Ali Cetin, Heather A. Sullivan, Bosiljka Tasic, Karel Svoboda, Julie Pendergraft, Osnat Penn, John W. Phillips, Ian R. Wickersham, Ed S. Lein, Loren L. Looger, Peter A. Groblewski, Hongkui Zeng, Allan R. Jones, Rachael Larsen, Emma Garren, Darren Bertagnolli, Michael N. Economo, Eliza Barkan, and Vilas Menon
- Subjects
0301 basic medicine ,Male ,Cell type ,Glutamic Acid ,Neocortex ,Biology ,Article ,Transcriptome ,03 medical and health sciences ,Glutamatergic ,Mice ,Single-cell analysis ,medicine ,Animals ,GABAergic Neurons ,Visual Cortex ,Multidisciplinary ,Sequence Analysis, RNA ,Gene Expression Profiling ,Motor Cortex ,Gene expression profiling ,030104 developmental biology ,medicine.anatomical_structure ,Visual cortex ,nervous system ,Organ Specificity ,Female ,Single-Cell Analysis ,Neuroscience ,Biomarkers ,Motor cortex - Abstract
The neocortex contains a multitude of cell types that are segregated into layers and functionally distinct areas. To investigate the diversity of cell types across the mouse neocortex, here we analysed 23,822 cells from two areas at distant poles of the mouse neocortex: the primary visual cortex and the anterior lateral motor cortex. We define 133 transcriptomic cell types by deep, single-cell RNA sequencing. Nearly all types of GABA (γ-aminobutyric acid)-containing neurons are shared across both areas, whereas most types of glutamatergic neurons were found in one of the two areas. By combining single-cell RNA sequencing and retrograde labelling, we match transcriptomic types of glutamatergic neurons to their long-range projection specificity. Our study establishes a combined transcriptomic and projectional taxonomy of cortical cell types from functionally distinct areas of the adult mouse cortex.
- Published
- 2017
35. A robot for high yield electrophysiology and morphology of single neurons in vivo
- Author
-
Lu Li, Christof Koch, William A. Stoy, Benjamin Ouellette, Craig R. Forest, Emma Garren, Tanya L. Daigle, and Hongkui Zeng
- Subjects
Male ,0301 basic medicine ,Science ,General Physics and Astronomy ,Hippocampal formation ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,Mice ,03 medical and health sciences ,In vivo ,Animals ,Neurons ,Multidisciplinary ,Electroporation ,Brain ,Equipment Design ,Robotics ,General Chemistry ,Anatomy ,Electrophysiological Phenomena ,Mice, Inbred C57BL ,Electrophysiology ,030104 developmental biology ,nervous system ,Models, Animal ,Robot ,Female ,Single-Cell Analysis ,Microelectrodes ,Neuroscience ,Software - Abstract
Single-cell characterization and perturbation of neurons provides knowledge critical to addressing fundamental neuroscience questions including the structure–function relationship and neuronal cell-type classification. Here we report a robot for efficiently performing in vivo single-cell experiments in deep brain tissues optically difficult to access. This robot automates blind (non-visually guided) single-cell electroporation (SCE) and extracellular electrophysiology, and can be used to characterize neuronal morphological and physiological properties of, and/or manipulate genetic/chemical contents via delivering extraneous materials (for example, genes) into single neurons in vivo. Tested in the mouse brain, our robot successfully reveals the full morphology of single-infragranular neurons recorded in multiple neocortical regions, as well as deep brain structures such as hippocampal CA3, with high efficiency. Our robot thus can greatly facilitate the study of in vivo full morphology and electrophysiology of single neurons in the brain., Single-cell characterization and perturbation of neurons is critical for revealing the structure-function relationship of brain cells. Here the authors develop a robot that performs single-cell electroporation and extracellular electrophysiology and can be used for performing in vivo single-cell experiments in deep brain tissues optically difficult to access.
- Published
- 2017
36. SmartScope2: Simultaneous Imaging and Reconstruction of Neuronal Morphology
- Author
-
Hongkui Zeng, Peter Saggau, Brian Long, Ed Lein, Ali Cetin, Michael Hawrylycz, Tanya L. Daigle, Jonathan T. Ting, Bosiljka Tasic, Zhi Zhou, Hanchuan Peng, and Ryder P. Gwinn
- Subjects
0301 basic medicine ,Computer science ,lcsh:Medicine ,Morphology (biology) ,Article ,Reduction (complexity) ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Imaging, Three-Dimensional ,Microscopy ,Animals ,Humans ,Computer vision ,lcsh:Science ,Automation, Laboratory ,Cerebral Cortex ,Neurons ,Multidisciplinary ,business.industry ,lcsh:R ,Pattern recognition ,Cortical neurons ,Automation ,030104 developmental biology ,Microscopy, Fluorescence, Multiphoton ,lcsh:Q ,Artificial intelligence ,business ,030217 neurology & neurosurgery - Abstract
Quantitative analysis of neuronal morphology is critical in cell type classification and for deciphering how structure gives rise to function in the brain. Most current approaches to imaging and tracing neuronal 3D morphology are data intensive. We introduce SmartScope2, the first open source, automated neuron reconstruction machine integrating online image analysis with automated multiphoton imaging. SmartScope2 takes advantage of a neuron’s sparse morphology to improve imaging speed and reduce image data stored, transferred and analyzed. We show that SmartScope2 is able to produce the complex 3D morphology of human and mouse cortical neurons with six-fold reduction in image data requirements and three times the imaging speed compared to conventional methods.
- Published
- 2017
37. Volumetric Ca2+ Imaging in the Mouse Brain Using Hybrid Multiplexed Sculpted Light Microscopy
- Author
-
Siegfried Weisenburger, Fraser T. Sparks, Tanya L. Daigle, Jason Manley, Francisca Martínez Traub, Alipasha Vaziri, Brandon Chen, Frank Tejera, Hongkui Zeng, Attila Losonczy, and Jeff Demas
- Subjects
Systems neuroscience ,0303 health sciences ,business.industry ,Posterior parietal cortex ,Hippocampus ,Biology ,Modular design ,Auditory cortex ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,Calcium imaging ,Microscopy ,medicine ,Neuron ,business ,030217 neurology & neurosurgery ,030304 developmental biology ,Biomedical engineering - Abstract
Calcium imaging using two-photon scanning microscopy has become an essential tool in neuroscience. However, in its typical implementation, the tradeoffs between fields of view, acquisition speeds, and depth restrictions in scattering brain tissue pose severe limitations. Here, using an integrated systems-wide optimization approach combined with multiple technical innovations, we introduce a new design paradigm for optical microscopy based on maximizing biological information while maintaining the fidelity of obtained neuron signals. Our modular design utilizes hybrid multi-photon acquisition and allows volumetric recording of neuroactivity at single-cell resolution within up to 1 × 1 × 1.22 mm volumes at up to 17 Hz in awake behaving mice. We establish the capabilities and potential of the different configurations of our imaging system at depth and across brain regions by applying it to in vivo recording of up to 12,000 neurons in mouse auditory cortex, posterior parietal cortex, and hippocampus.
- Published
- 2019
38. In vivo sub-millisecond two-photon optogenetics with temporally focused patterned light
- Author
-
Deniz Dalkara, Valentina Emiliani, Tanya L. Daigle, Emiliano Ronzitti, Brian Lee, I-Wen Chen, Hongkui Zeng, Eirini Papagiakoumou, Chen, I-Wen, Appel à projets générique - Éclairer la fonction des neurones hubs de l'hippocampe adulte chez la souris éveillée par microscopie à modulation de front d'onde - - HOLOHUB2014 - ANR-14-CE13-0016 - Appel à projets générique - VALID, Holographie tri-dimensionelle pour l'activation parallèle des neurones - - 3DHoloPAc2015 - ANR-15-CE19-0001 - AAPG2015 - VALID, Holistic evaluation of light and multiwave applications to high resolution imaging in ophthalmic translational research revisiting the helmholtzian synergies - HELMHOLTZ - - EC:FP7:ERC2014-08-01 - 2020-07-31 - 610110 - VALID, Institut de la Vision, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Neurophotonique (UMR 8250), Université Paris Descartes - Paris 5 (UPD5)-Centre National de la Recherche Scientifique (CNRS), Allen Institute for Brain Science [Seattle, WA, USA], This work was supported by the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie Grant Agreement no. 747598 (to I.-W.C.). E.P. and V.E. acknowledge support from the Agence Nationale de la Recherche ANR (Grants ANR-14-CE13-0016, Holohub and ANR-15-CE19-0001-01, 3DHoloPAc). V.E. acknowledges support from the Human Frontiers Science Program (Grant RGP0015/2016), the National Institutes of Health (Grant NIH U01NS090501-03), and Gettylab. E.R. received funding from the European Research Council Synergy Grant scheme (Helmholtz, ERC Grant Agreement 610110)., ANR-14-CE13-0016,HOLOHUB,Éclairer la fonction des neurones hubs de l'hippocampe adulte chez la souris éveillée par microscopie à modulation de front d'onde(2014), ANR-15-CE19-0001,3DHoloPAc,Holographie tri-dimensionelle pour l'activation parallèle des neurones(2015), European Project: 610110,EC:FP7:ERC,ERC-2013-SyG,HELMHOLTZ(2014), and Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)
- Subjects
0301 basic medicine ,computer-generated holography ,millisecond photoactivation ,Optogenetics ,in vivo two-photon optogenetics ,Cell morphology ,03 medical and health sciences ,0302 clinical medicine ,Calcium imaging ,Two-photon excitation microscopy ,medicine ,Premovement neuronal activity ,[SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] ,Research Articles ,Physics ,General Neuroscience ,Computer-generated holography ,030104 developmental biology ,Visual cortex ,medicine.anatomical_structure ,Temporal resolution ,temporal focusing ,mouse visual cortex ,[SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC] ,Neuroscience ,030217 neurology & neurosurgery - Abstract
To better examine circuit mechanisms underlying perception and behavior, researchers need tools to enable temporally precise control of action-potential generation of individual cells from neuronal ensembles. Here we demonstrate that such precision can be achieved with two-photon (2P) temporally focused computer-generated holography to control neuronal excitability at the supragranular layers of anesthetized and awake visual cortex in both male and female mice. Using 2P-guided whole-cell or cell-attached recordings in positive neurons expressing any of the three opsins ReaChR, CoChR, or ChrimsonR, we investigated the dependence of spiking activity on the opsin's channel kinetics. We found that in all cases the use of brief illumination (≤10 ms) induces spikes of millisecond temporal resolution and submillisecond precision, which were preserved upon repetitive illuminations up to tens of hertz. To reach high temporal precision, we used a large illumination spot covering the entire cell body and an amplified laser at high peak power and low excitation intensity (on average ≤0.2 mW/μm(2)), thus minimizing the risk for nonlinear photodamage effects. Finally, by combining 2P holographic excitation with electrophysiological recordings and calcium imaging using GCaMP6s, we investigated the factors, including illumination shape and intensity, opsin distribution in the target cell, and cell morphology, which affect the spatial selectivity of single-cell and multicell holographic activation. Parallel optical control of neuronal activity with cellular resolution and millisecond temporal precision should make it easier to investigate neuronal connections and find further links between connectivity, microcircuit dynamics, and brain functions. SIGNIFICANCE STATEMENT Recent developments in the field of optogenetics has enabled researchers to probe the neuronal microcircuit with light by optically actuating genetically encoded light-sensitive opsins expressed in the target cells. Here, we applied holographic light shaping and temporal focusing to simultaneously deliver axially confined holographic patterns to opsin-positive cells in the living mouse cortex. Parallel illumination efficiently induced action potentials with high temporal resolution and precision for three opsins of different kinetics. We extended the parallel optogenetic activation at low intensity to multiple neurons and concurrently monitored their calcium dynamics. These results demonstrate fast and temporally precise in vivo control of a neuronal subpopulation, opening new opportunities for revealing circuit mechanisms underlying brain functions.
- Published
- 2019
39. Elimination of GRK2 from Cholinergic Neurons Reduces Behavioral Sensitivity to Muscarinic Receptor Activation
- Author
-
Marc G. Caron and Tanya L. Daigle
- Subjects
Pain Threshold ,medicine.medical_specialty ,G-Protein-Coupled Receptor Kinase 2 ,Mice, Transgenic ,Motor Activity ,Muscarinic Agonists ,Biology ,Article ,Body Temperature ,Choline O-Acetyltransferase ,Mice ,Cocaine ,Dopamine Uptake Inhibitors ,Internal medicine ,Muscarinic acetylcholine receptor ,Muscarinic acetylcholine receptor M5 ,medicine ,Muscarinic acetylcholine receptor M4 ,Oxotremorine ,Animals ,Cholinergic neuron ,Analysis of Variance ,General Neuroscience ,Brain ,Muscarinic acetylcholine receptor M3 ,Muscarinic acetylcholine receptor M2 ,Receptors, Muscarinic ,Cholinergic Neurons ,Conditioned place preference ,Mice, Inbred C57BL ,Endocrinology ,Hyperalgesia ,Conditioning, Operant ,Salivation ,Neuroscience ,medicine.drug - Abstract
Although G-protein-coupled receptor kinase 2 (GRK2) is the most widely studied member of a family of kinases that has been shown to exert powerful influences on a variety of G-protein-coupled receptors, its role in the brain remains largely unknown. Here we report the localization of GRK2 in the mouse brain and generate novel conditional knock-out (KO) mice to assess the physiological importance of this kinase in cholinergic neurons. Mice with the selective deletion of GRK2 in this cell population (ChATIRES-creGrk2f/fKO mice) exhibit reduced behavioral responsiveness to challenge with oxotremorine-M (Oxo-M), a nonselective muscarinic acetylcholine receptor agonist. Specifically, Oxo-M-induced hypothermia, hypolocomotion, and salivation were markedly reduced in these animals, while analgesic responses were unaltered. In contrast, we found that GRK2 deficiency in cholinergic neurons does not alter cocaine-induced psychomotor activation, behavioral sensitization, or conditioned place preference. These results demonstrate that the elimination of GRK2 in cholinergic neurons reduces sensitivity to select muscarinic-mediated behaviors, while dopaminergic effects remain intact and further suggests that GRK2 may selectively impair muscarinic acetylcholine receptor-mediated functionin vivo.
- Published
- 2012
40. Opposite function of dopamine D1 and N-methyl-D-aspartate receptors in striatal cannabinoid-mediated signaling
- Author
-
William C. Wetsel, Tanya L. Daigle, and Marc G. Caron
- Subjects
AM251 ,medicine.medical_specialty ,Cannabinoid receptor ,Beta-Arrestins ,General Neuroscience ,medicine.medical_treatment ,Biology ,Pharmacology ,Endocrinology ,Dopamine receptor D1 ,Dopamine receptor D2 ,Internal medicine ,medicine ,Cannabinoid receptor type 2 ,Cannabinoid receptor antagonist ,Cannabinoid ,medicine.drug - Abstract
It is well established that the cannabinoid and dopamine systems interact at various levels to regulate basal ganglia function. Although it is well known that acute administration of cannabinoids to mice can modify dopamine-dependent behaviors, the intraneuronal signaling pathways employed by these agents in the striatum are not well understood. Here we used knockout mouse models to examine the regulation of striatal extracellular-signal-regulated kinases 1 and 2 (ERK1/2) signaling by behaviorally relevant doses of cannabinoids. This cellular pathway has been implicated as a central mediator of drug reward and synaptic plasticity. In C57BL/6J mice, acute administration of the cannabinoid agonists, (-)-11-hydroxydimethylheptyl-Δ8-tetrahydrocannabinol (HU-210) and delta-9-tetrahydrocannabinol (Δ(9) -THC), promoted a dose- and time-dependent decrease in the phosphorylation of ERK1/2 in dorsal striatum. Co-administration of the CB1 cannabinoid receptor antagonist N-(Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide(AM251) with HU-210 prevented ERK1/2 inactivation, indicating a requirement for activation of this receptor. In dopamine D1 receptor knockout animals treated with HU-210, the magnitude of the HU-210-dependent decrease in striatal ERK1/2 signaling was greater than in wild-type controls. In contrast, HU-210 administration to N-methyl-D-aspartate receptor knockdown mice was ineffective at promoting striatal ERK1/2 inactivation. Genetic deletion of other potential ERK1/2 mediators, the dopamine D2 receptors or β-arrestin-1 or -2, did not affect the HU-210-induced modulation of ERK1/2 signaling in the striatum. These results support the hypothesis that dopamine D1 receptors and N-methyl-D-aspartate receptors act in an opposite manner to regulate striatal CB1 cannabinoid receptor signal transduction.
- Published
- 2011
41. A Dopamine D1 Receptor-Dependent β-Arrestin Signaling Complex Potentially Regulates Morphine-Induced Psychomotor Activation but not Reward in Mice
- Author
-
Nikhil M. Urs, Marc G. Caron, and Tanya L. Daigle
- Subjects
Male ,Narcotics ,Time Factors ,Arrestins ,Motor Activity ,Pharmacology ,Biology ,Mice ,Dopamine receptor D1 ,Reward ,Dopamine ,Arrestin ,medicine ,Animals ,Immunoprecipitation ,Enzyme Inhibitors ,Extracellular Signal-Regulated MAP Kinases ,Receptor ,beta-Arrestins ,Mice, Knockout ,Analysis of Variance ,Dopamine Plasma Membrane Transport Proteins ,Behavior, Animal ,Morphine ,Beta-Arrestins ,Receptors, Dopamine D1 ,beta-Arrestin 2 ,Conditioned place preference ,Mice, Inbred C57BL ,Psychiatry and Mental health ,Gene Expression Regulation ,Dopamine receptor ,Conditioning, Operant ,Original Article ,Female ,Signal transduction ,Psychomotor Performance ,Signal Transduction ,medicine.drug - Abstract
Morphine is a widely used analgesic in humans that is associated with multiple untoward effects, such as addiction and physical dependence. In rodent models, morphine also induces locomotor activity. These effects likely involve functionally selective mechanisms. Indeed, G protein-coupled receptor desensitization and adaptor protein β-arrestin 2 (βarr2) through its interaction with the μ-opioid receptor regulates the analgesic but not the rewarding properties of morphine. However, βarr2 is also required for morphine-induced locomotor activity in mice, but the exact cellular and molecular mechanisms that mediate this arrestin-dependent behavior are not understood. In this study, we show that βarr2 is required for morphine-induced locomotor activity in a dopamine D1 receptor (D1R)-dependent manner and that a βarr2/phospho-ERK (βarr2/pERK) signaling complex may mediate this behavior. Systemic administration of SL327, an MEK inhibitor, inhibits morphine-induced locomotion in wild-type mice in a dose-dependent manner. Acute morphine administration to mice promotes the formation of a βarr2/pERK signaling complex. Morphine-induced locomotor activity and formation of the βarr2/pERK signaling complex is blunted in D1R knockout (D1-KO) mice and is presumably independent of D2 dopamine receptors. However, D1Rs are not required for morphine-induced reward as D1-KO mice show the same conditioned place preference for morphine as do control mice. Taken together, these results suggest a potential role for a D1R-dependent βarr2/pERK signaling complex in selectively mediating the locomotor-stimulating but not the rewarding properties of morphine.
- Published
- 2010
42. A Suite of Transgenic Driver and Reporter Mouse Lines with Enhanced Brain-Cell-Type Targeting and Functionality
- Author
-
Travis A. Hage, Christopher A. Baker, Linda Madisen, Alice Bosma-Moody, Rylan S. Larsen, Matthew T. Valley, Jonathan T. Ting, Karla E. Hirokawa, Kimberly A. Smith, Olivia Fong, Jérôme Lecoq, Garreck H. Lenz, Julie Pendergraft, Susan M. Sunkin, Julie A. Harris, La'Akea Siverts, Maya Mills, Zizhen Yao, Michael Z. Lin, Thuc Nghi Nguyen, Mariya Chavarha, Philip R. Nicovich, Nuno Maçarico da Costa, Lawrence Huang, Lu Li, Miranda Walker, Marc Takeno, Gabe J. Murphy, Lucas T. Graybuck, Jack Waters, Emma Garren, Edward S. Boyden, Medea McGraw, Rachael Larsen, James Harrington, Douglas R. Ollerenshaw, Ulf Knoblich, Tanya L. Daigle, Hongkui Zeng, Bosiljka Tasic, and Hong Gu
- Subjects
0301 basic medicine ,Genetically modified mouse ,Cell type ,RNA, Untranslated ,Light ,Transgene ,Cell ,Mice, Transgenic ,Computational biology ,Optogenetics ,Biology ,Brain Cell ,General Biochemistry, Genetics and Molecular Biology ,Cell Line ,Gene Knockout Techniques ,Mice ,03 medical and health sciences ,Genes, Reporter ,health services administration ,medicine ,Animals ,natural sciences ,Transgenes ,In Situ Hybridization, Fluorescence ,Neurons ,Brain ,Transgenesis ,030104 developmental biology ,medicine.anatomical_structure ,Microscopy, Fluorescence ,Calcium ,human activities ,Function (biology) - Abstract
Modern genetic approaches are powerful in providing access to diverse cell types in the brain and facilitating the study of their function. Here, we report a large set of driver and reporter transgenic mouse lines, including 23 new driver lines targeting a variety of cortical and subcortical cell populations and 26 new reporter lines expressing an array of molecular tools. In particular, we describe the TIGRE2.0 transgenic platform and introduce Cre-dependent reporter lines that enable optical physiology, optogenetics, and sparse labeling of genetically defined cell populations. TIGRE2.0 reporters broke the barrier in transgene expression level of single-copy targeted-insertion transgenesis in a wide range of neuronal types, along with additional advantage of a simplified breeding strategy compared to our first-generation TIGRE lines. These novel transgenic lines greatly expand the repertoire of high-precision genetic tools available to effectively identify, monitor, and manipulate distinct cell types in the mouse brain.
- Published
- 2018
43. Regulation of CB1cannabinoid receptor internalization by a promiscuous phosphorylation-dependent mechanism
- Author
-
Ken Mackie, Tanya L. Daigle, and Mary Lee Kwok
- Subjects
Cannabinoid receptor ,Arrestins ,media_common.quotation_subject ,education ,Biology ,Endocytosis ,Biochemistry ,Article ,Mice ,Cellular and Molecular Neuroscience ,Receptor, Cannabinoid, CB1 ,Arrestin ,Animals ,Humans ,Amino Acid Sequence ,Phosphorylation ,Receptor ,Internalization ,health care economics and organizations ,beta-Arrestins ,media_common ,G protein-coupled receptor ,Cannabinoids ,Beta-Arrestins ,Cell Membrane ,beta-Arrestin 2 ,Protein Structure, Tertiary ,Cell biology ,Molecular Weight ,Protein Transport - Abstract
Agonists stimulate CB1 receptor internalization. Previous work suggests that the extreme carboxy-terminus of the receptor regulates this internalization – likely through the phosphorylation of serines and threonines clustered within this region. While truncation of the carboxy-terminus (V460Z CB1) and consequent removal of these putative phosphorylation sites prevents endocytosis in AtT20 cells, the residues necessary for CB1 receptor internalization remain elusive. To determine the structural requirements for internalization, we evaluated endocytosis of carboxy-terminal mutant CB1 receptors stably expressed in HEK293 cells. In contrast to AtT20 cells, V460Z CB1 receptors expressed in HEK293 cells internalized to the same extent and with similar kinetics as the wild-type receptor. However, mutation of serine and/or threonine residues within the extreme carboxy-terminal attenuated internalization when these receptors were expressed in HEK293 cells. These results establish that the extreme carboxy-terminal phosphorylation sites are not required for internalization of truncated receptors, but are required for internalization of full-length receptors in HEK293 cells. Analysis of βarrestin-2 recruitment to mutant CB1 receptors suggests that putative carboxy-terminal phosphorylation sites mediate βarrestin-2 translocation. This study indicates that the local cellular environment affects the structural determinants of CB1 receptor internalization. Additionally, phosphorylation likely regulates the internalization of (full-length) CB1 receptors.
- Published
- 2008
44. Targeting β-arrestin2 in the treatment of <scp>l</scp> -DOPA–induced dyskinesia in Parkinson’s disease
- Author
-
Erwan Bezard, Simone Bido, Raul R. Gainetdinov, Sean M. Peterson, Tanya L. Daigle, Caroline E. Bass, Marc G. Caron, and Nikhil M. Urs
- Subjects
Male ,Dyskinesia, Drug-Induced ,Levodopa ,Parkinson's disease ,Arrestins ,Dopamine ,Pharmacology ,Rats, Sprague-Dawley ,Mice ,Dopamine receptor D2 ,medicine ,Animals ,Oxidopamine ,beta-Arrestins ,Mice, Knockout ,Neurons ,Dyskinesias ,Multidisciplinary ,Behavior, Animal ,business.industry ,Beta-Arrestins ,Parkinson Disease ,Viral rescue ,medicine.disease ,nervous system diseases ,Rats ,Up-Regulation ,Mice, Inbred C57BL ,Disease Models, Animal ,PNAS Plus ,Dyskinesia ,1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine ,Dopamine receptor ,Macaca ,medicine.symptom ,business ,Gene Deletion ,Signal Transduction ,medicine.drug - Abstract
Parkinson's disease (PD) is characterized by severe locomotor deficits and is commonly treated with the dopamine (DA) precursor l-3,4-dihydroxyphenylalanine (L-DOPA), but its prolonged use causes dyskinesias referred to as L-DOPA-induced dyskinesias (LIDs). Recent studies in animal models of PD have suggested that dyskinesias are associated with the overactivation of G protein-mediated signaling through DA receptors. β-Arrestins desensitize G protein signaling at DA receptors (D1R and D2R) in addition to activating their own G protein-independent signaling events, which have been shown to mediate locomotion. Therefore, targeting β-arrestins in PD L-DOPA therapy might prove to be a desirable approach. Here we show in a bilateral DA-depletion mouse model of Parkinson's symptoms that genetic deletion of β-arrestin2 significantly limits the beneficial locomotor effects while markedly enhancing the dyskinesia-like effects of acute or chronic L-DOPA treatment. Viral rescue or overexpression of β-arrestin2 in knockout or control mice either reverses or protects against LIDs and its key biochemical markers. In other more conventional animal models of DA neuron loss and PD, such as 6-hydroxydopamine-treated mice or rats and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated nonhuman primates, β-arrestin2 overexpression significantly reduced dyskinesias while maintaining the therapeutic effect of L-DOPA. Considerable efforts are being spent in the pharmaceutical industry to identify therapeutic approaches to block LIDs in patients with PD. Our results point to a potential therapeutic approach, whereby development of either a genetic or pharmacological intervention to enhance β-arrestin2- or limit G protein-dependent D1/D2R signaling could represent a more mechanistically informed strategy.
- Published
- 2015
45. Relevance of GPCR functional selectivity/biased signaling to drugs of abuse (468.1)
- Author
-
Marc G. Caron, Nikhil M. Urs, Sean M. Peterson, Joshua C. Snyder, and Tanya L. Daigle
- Subjects
Drugs of abuse ,Cell signaling ,G protein ,Chemistry ,fungi ,food and beverages ,Pharmacology ,Biochemistry ,Genetics ,Functional selectivity ,Molecular Biology ,Neuroscience ,Intracellular ,Biotechnology ,G protein-coupled receptor - Abstract
We now understand that GPCRs can signal not only through activation of G proteins but also through the ability of β-arrestins to scaffold intracellular signaling molecules with distinct temporal an...
- Published
- 2014
46. Acute Brain Slice Methods for Adult and Aging Animals: Application of Targeted Patch Clamp Analysis and Optogenetics
- Author
-
Guoping Feng, Jonathan T. Ting, Qian Chen, and Tanya L. Daigle
- Subjects
Neurons ,Genetically modified mouse ,Aging ,Histocytological Preparation Techniques ,Patch-Clamp Techniques ,Transgene ,Brain ,Gene Expression ,Channelrhodopsin ,Mice, Transgenic ,Equipment Design ,Biology ,Optogenetics ,Article ,Mice ,Synaptic function ,Slice preparation ,Channelrhodopsins ,Animals ,Premovement neuronal activity ,Patch clamp ,Neuroscience - Abstract
The development of the living acute brain slice preparation for analyzing synaptic function roughly a half century ago was a pivotal achievement that greatly influenced the landscape of modern neuroscience. Indeed, many neuroscientists regard brain slices as the gold-standard model system for detailed cellular, molecular, and circuitry level analysis and perturbation of neuronal function. A critical limitation of this model system is the difficulty in preparing slices from adult and aging animals, and over the past several decades few substantial methodological improvements have emerged to facilitate patch clamp analysis in the mature adult stage. In this chapter we describe a robust and practical protocol for preparing brain slices from mature adult mice that are suitable for patch clamp analysis. This method reduces swelling and damage in superficial layers of the slices and improves the success rate for targeted patch clamp recordings, including recordings from fluorescently labeled populations in slices derived from transgenic mice. This adult brain slice method is suitable for diverse experimental applications, including both monitoring and manipulating neuronal activity with genetically encoded calcium indicators and optogenetic actuators, respectively. We describe the application of this adult brain slice platform and associated methods for screening kinetic properties of Channelrhodopsin (ChR) variants expressed in genetically-defined neuronal subtypes.
- Published
- 2014
47. Targeting Beta-Arrestin Dependent Signaling in the Treatment of Parkinson’s Disease
- Author
-
Tanya L. Daigle, Jonathan T. Ting, Nikhil M. Urs, and Marc G. Caron
- Subjects
Parkinson's disease ,business.industry ,Cancer research ,Medicine ,Arrestin beta 2 ,business ,medicine.disease - Published
- 2014
48. Selective deletion of GRK2 alters psychostimulant-induced behaviors and dopamine neurotransmission
- Author
-
Mark J. Ferris, Marc G. Caron, Tatyana D. Sotnikova, Nikhil M. Urs, Tanya L. Daigle, Raul R. Gainetdinov, and Sara R. Jones
- Subjects
medicine.medical_specialty ,G-Protein-Coupled Receptor Kinase 2 ,Receptor, Adenosine A2A ,Dopamine Plasma Membrane Transport Proteins ,Dopamine ,Biology ,Neurotransmission ,Motor Activity ,Synaptic Transmission ,Dopamine receptor D1 ,Cocaine ,Internal medicine ,Dopamine receptor D2 ,Conditional gene knockout ,Conditioning, Psychological ,medicine ,Animals ,Pharmacology ,Mice, Knockout ,Neurons ,Receptors, Dopamine D2 ,Beta adrenergic receptor kinase ,Receptors, Dopamine D1 ,Brain ,Psychiatry and Mental health ,Endocrinology ,Space Perception ,Autoreceptor ,biology.protein ,Central Nervous System Stimulants ,Original Article ,medicine.drug - Abstract
GRK2 is a G protein-coupled receptor kinase (GRK) that is broadly expressed and is known to regulate diverse types of receptors. GRK2 null animals exhibit embryonic lethality due to a severe developmental heart defect, which has precluded the study of this kinase in the adult brain. To elucidate the specific role of GRK2 in the brain dopamine (DA) system, we used a conditional gene knockout approach to selectively delete GRK2 in DA D1 receptor (D1R)-, DA D2 receptor (D2R)-, adenosine 2A receptor (A2AR)-, or DA transporter (DAT)-expressing neurons. Here we show that select GRK2-deficient mice display hyperactivity, hyposensitivity, or hypersensitivity to the psychomotor effects of cocaine, altered striatal signaling, and DA release and uptake. Mice with GRK2 deficiency in D2R-expressing neurons also exhibited increased D2 autoreceptor activity. These findings reveal a cell-type-specific role for GRK2 in the regulation of normal motor behavior, sensitivity to psychostimulants, dopamine neurotransmission, and D2 autoreceptor function.
- Published
- 2013
49. N-Aryl Piperazine Metabotropic Glutamate Receptor 5 Positive Allosteric Modulators Possess Efficacy in Preclinical Models of NMDA Hypofunction and Cognitive Enhancement
- Author
-
J.T. Manka, H. Lavreysen, Colleen M. Niswender, A. Ahnaou, Shaun R. Stauffer, Carrie K. Jones, Marc G. Caron, Tanya L. Daigle, Thomas M. Bridges, B.J. Hrupka, E.J. Herman, Charles David Weaver, Wilhelmus Drinkenburg, Amy J. Ramsey, A.S. Hammond, Gregor James Macdonald, Claire Mackie, Thomas Steckler, Satyawan Jadhav, Craig W. Lindsley, Karen J. Gregory, José M. Bartolomé, P.J. Conn, and Nellie Byun
- Subjects
Male ,Psychosis ,Receptor, Metabotropic Glutamate 5 ,Drug Evaluation, Preclinical ,Pharmacology ,Hyperkinesis ,Motor Activity ,Transfection ,Receptors, N-Methyl-D-Aspartate ,Piperazines ,Rats, Sprague-Dawley ,Glutamatergic ,Mice ,Neuropharmacology ,Allosteric Regulation ,medicine ,Animals ,Humans ,Maze Learning ,Prepulse inhibition ,Nootropic Agents ,Mice, Knockout ,Dose-Response Relationship, Drug ,Metabotropic glutamate receptor 5 ,Glutamate receptor ,medicine.disease ,Rats ,HEK293 Cells ,Memory, Short-Term ,Metabotropic glutamate receptor ,Schizophrenia ,Molecular Medicine ,NMDA receptor ,Psychology ,Neuroscience ,Ionotropic effect ,Antipsychotic Agents - Abstract
Impaired transmission through glutamatergic circuits has been postulated to play a role in the underlying pathophysiology of schizophrenia. Furthermore, inhibition of the N-methyl-d-aspartate (NMDA) subtype of ionotropic glutamate receptors (NMDAR) induces a syndrome that recapitulates many of the symptoms observed in patients with schizophrenia. Selective activation of metabotropic glutamate receptor subtype 5 (mGlu5) may provide a novel therapeutic approach for treatment of symptoms associated with schizophrenia through facilitation of transmission through central glutamatergic circuits. Here, we describe the characterization of two novel N-aryl piperazine mGlu5 positive allosteric modulators (PAMs): 2-(4-(2-(benzyloxy)acetyl)piperazin-1-yl)benzonitrile (VU0364289) and 1-(4-(2,4-difluorophenyl)piperazin-1-yl)-2-((4-fluorobenzyl)oxy)ethanone (DPFE). VU0364289 and DPFE induced robust leftward shifts in the glutamate concentration-response curves for Ca(2+) mobilization and extracellular signal-regulated kinases 1 and 2 phosphorylation. Both PAMs displayed micromolar affinity for the common mGlu5 allosteric binding site and high selectivity for mGlu5. VU0364289 and DPFE possessed suitable pharmacokinetic properties for dosing in vivo and produced robust dose-related effects in reversing amphetamine-induced hyperlocomotion, a preclinical model predictive of antipsychotic-like activity. In addition, DPFE enhanced acquisition of contextual fear conditioning in rats and reversed behavioral deficits in a mouse model of NMDAR hypofunction. In contrast, DPFE had no effect on reversing apomorphine-induced disruptions of prepulse inhibition of the acoustic startle reflex. These mGlu5 PAMs also increased monoamine levels in the prefrontal cortex, enhanced performance in a hippocampal-mediated memory task, and elicited changes in electroencephalogram dynamics commensurate with procognitive effects. Collectively, these data support and extend the role for the development of novel mGlu5 PAMs for the treatment of psychosis and cognitive deficits observed in individuals with schizophrenia.
- Published
- 2013
50. Protocol Development for 3D Reconstructions: Combining In Vivo Imaging, APEX2 and En Bloc Staining of Mouse Visual Cortex
- Author
-
Adam Bleckert, Ali Cetin, Derrick Brittain, Agnes L. Bodor, Marc Takeno, Daniel J. Bumbarger, JoAnn Buchanan, Tanya L. Daigle, and Nuno Maçarico da Costa
- Subjects
Pathology ,medicine.medical_specialty ,Visual cortex ,medicine.anatomical_structure ,Chemistry ,medicine ,Instrumentation ,Preclinical imaging ,Staining - Published
- 2015
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.